CN108472649A

CN108472649A - The microfluidic device and related systems and methods on the electrowetting surface with optimization

Info

Publication number: CN108472649A
Application number: CN201680077445.7A
Authority: CN
Inventors: R·D·小罗威; S·N·裴; J·宫; A·J·马斯楚安尼; J·M·麦克尤恩; J·K·瓦利
Original assignee: Berkeley's Light Life Technology Co
Current assignee: Berkeley's Light Life Technology Co
Priority date: 2015-10-27
Filing date: 2016-10-27
Publication date: 2018-08-31
Anticipated expiration: 2036-10-27
Also published as: IL258851B2; TW202126571A; TWI781484B; TWI714656B; AU2021286430A1; IL258851B1; CA3001616A1; IL258851A; CA3001616C; WO2017075295A8; WO2017075295A1; JP2018535088A; KR20180072812A; EP3862088A1; SG10202107069UA; CN114289087B; JP2021126656A; KR102426825B1; SG11201802968VA; JP6891169B2

Abstract

Provide the microfluidic device with electrowetting configuration and the drop actuation surfaces optimized.The equipment includes the conductive substrates for having dielectric layer, be covalently bound to the hydrophobic layer of dielectric layer and be electrically coupled to dielectric layer and be configured for connection to the first electrode of voltage source.Microfluidic device further includes the second electrode being optionally included in lid, is configured for connection to voltage source.Hydrophobic layer is characterized in that being covalently bound to the self-association molecule of dielectric layer surface in a manner of generating dense accumulation single layer, and the single layer resists the insertion and/or infiltration of polar molecule or substance.It additionally provides：Microfluidic device has electrowetting configuration, further comprises the part that there is dielectrophoresis to configure or module；System, including combination have any microfluidic device of aqueous drop and the fluid media (medium) unmixing with the medium of aqueous drop；Related kit；And the method that drop is manipulated in microfluidic device, drop include optionally microorganism, such as biological cell.

Description

The microfluidic device and related systems and methods on the electrowetting surface with optimization

Cross reference to related applications

This application claims No. 62/246,605 U.S. Provisional Application submitted on October 27th, 2015, October 28 in 2015 No. 62/342,131 U.S. that No. 62/247,725 U.S. Provisional Application of day submission, on May 26th, 2016 submit is interim The content of the priority of application and the 62/410th, No. 238 U.S. Provisional Application submitted on October 19th, 2016, each of which is logical Reference is crossed to be integrally incorporated herein.The application is the portion for No. 15/135,707 U.S. Patent application submitted on April 22nd, 2016 Continuation application, content is divided to be incorporated herein by reference in their entirety.

Background technology

Speck body (such as biological cell) can be handled in microfluidic devices.For example, including speck body or reagent Drop can move and merge in microfluidic device.The embodiment of the present invention is related to the improvement to microfluidic device, promotees Into the robustness manipulation to drop, allows accurate on small scale where and reproducibly execute complicated chemistry and biological respinse.It is logical The effective wetting property for changing the electrowetting surface in microfluidic device is crossed, drop can be made to move and close in microfluidic device And.This movement can promote to handle the workflow of cell, optionally to be assessed after culture cell in microfluidic device Various cellularities.Current solution for electrowetting is substantially extremely limited, and can not extend or realize Additional function.Stable substrate therefore, it is necessary to improved electrowetting surface, for microfluidic applications and integration additional function (such as made it possible by electrowetting cell growth and characterization) before downstream processes, it is all these to will be helpful to others Medical research application.

Invention content

In one aspect, the present invention provides the microfluidic devices configured including electrowetting comprising has drop actuating The substrate on surface, drop actuation surfaces include the hydrophobic layer on the surface for being covalently bound to following dielectric layer (that is, inner-dielectric-ayer) (that is, outer hydrophobic layer) (or be made from it or consisting essentially of).When microfluidic device is operably connected to voltage source, It is placed on hydrophobic layer or otherwise the aqueous drop of contact hydrophobic layer can be reliable by electrowetting power and be robustly moistened It is wet and thus move.

Microfluidic device may include include the base portion of substrate, and substrate can further have be configured to connect to At least one electrode (for example, first electrode) of voltage source (for example, AC voltage sources), at least one electrode are electrically coupled to interior Jie Electric layer.In some embodiments, microfluidic device further includes lid and at least one spacer element.Substrate and lid can substantially each other It is parallel and be bonded together by spacer element with limit be configured to keep liquid medium capsule.In such embodiment In, lid may include at least one electrode for being configured to connect to voltage source (for example, AC voltage sources).In some embodiments In, microfluidic device may include unilateral electrowetting configuration.In such embodiments, microfluidic device need not include lid. For example, base portion may include substrate and be configured to connect to the first electrode of voltage source (such as AC voltage sources), and substrate It may include the second electrode (for example, mesh electrode) for being configured to connect to voltage source.

In some embodiments, outer hydrophobic layer includes being covalently bond to the self-association molecule of inner-dielectric-ayer to form fine and close heap Long-pending hydrophobic monolayer.In some embodiments, the self-association molecule of hydrophobic monolayer includes each siloxane group.In other implementations In example, the self-association molecule of hydrophobic monolayer includes each phosphonyl group.Siloxane group or phosphonyl group can be covalently bound to The surface of inner-dielectric-ayer.In some embodiments, the self-association molecule of hydrophobic monolayer includes each surface modified ligand and by table Face modified ligand directly or indirectly links to the link group on inner-dielectric-ayer surface.Surface modified ligand can be disclosed herein Any surface modified ligand.For example, surface modified ligand may include aliphatic group, such as alkane group.Thus, for example, dredging The self-association molecule of water monolayer can be alkyl-blocked siloxanes or alkyl-blocked Phosphonic acid molecules.Alkyl may include at least 10 carbon (for example, at least 14,16,18,20,22 or more carbon) chain (such as non-branched).In other embodiments, surface Modified ligand may include fluorine-substituted aliphatic group, such as fluoroalkyl.Thus, for example, self-association molecule can be fluoroalkyl The Phosphonic acid molecules of siloxanes or the fluoroalkyl sealing end of sealing end.Fluoroalkyl may include at least ten carbon (for example, at least 14,16, 18,20,22 or more carbon) chain (for example, non-branched).In certain embodiments, fluoroalkyl includes one or more (examples Such as, at least 4,6,8,10,12 or more) perfluocarbon.For example, fluoro-alkyl can have chemical formula CF₃-(CF₂)m- (CH₂) n-, wherein m is at least 2, n and is at least 2, and m+n is at least 9.In some embodiments, surface modified ligand includes Ehter bond between first aliphatic group and the second aliphatic group.For example, the first aliphatic group can be alkyl, the second aliphatic group Can be fluoroalkyl (for example, perfluoroalkyl).In some embodiments, the alkyl of surface modified ligand or fluoroalkyl are no branches 's.In some embodiments, the alkyl of surface modified ligand or fluoroalkyl are free of any cyclic structure.

In some embodiments, the outer hydrophobic layer of substrate has the thickness less than 5 nanometers (for example, about 1.5 to 3.0 nanometers) Degree.In some embodiments, the outer hydrophobic layer of substrate can be patterned such that compared with the rest part of outer hydrophobic layer, choosing Select region relative hydropathic.

In some embodiments, the inner-dielectric-ayer of substrate may include first layer dielectric material.For example, inner-dielectric-ayer can be with It is made of single layer dielectric material.First layer dielectric material may include oxide, such as metal oxide layer (such as aluminium oxide, Hafnium oxide etc.).In certain embodiments, the first oxide skin(coating) is formed by atomic layer deposition (ALD).Alternatively, inner-dielectric-ayer can To be the dielectric stack for including two or more layers dielectric material.Therefore, in certain embodiments, inner-dielectric-ayer may include First layer dielectric material and second layer dielectric material.First layer dielectric material may include oxide, such as metal oxide (example Such as aluminium oxide, hafnium oxide)；And second layer dielectric material may include the oxide or such as silicon nitride of such as silica Nitride.In such embodiments, first layer dielectric material can have contact second layer dielectric material first surface and With the covalently bound apparent surface of hydrophobic layer.In certain embodiments, the type of used dielectric material, the second layer are depended on Dielectric material can be with the thickness of about 30nm to about 100nm.For example, second layer dielectric material can include silica and can be with Thickness with about 30nm to about 50nm or about 30nm to about 40nm.Optionally, second layer dielectric material may include silicon nitride And it can be with the thickness of about 50nm to about 100nm or about 80nm to about 100nm.In certain embodiments, the second layer is situated between Electric material is formed by ALD.In other embodiments, second layer dielectric material is by plasma enhanced chemical vapor deposition (PECVD) technology is formed.In certain embodiments, first layer dielectric material can with about 10nm to about 50nm thickness (for example, About 10nm to about 20nm, about 15nm to about 25nm, about 20nm to about 30nm, about 25nm to about 35nm, about 30nm to about 40nm, about Any range that 35nm to about 45nm, about 40nm are limited to about 50nm or by two aforementioned endpoints), and ALD shapes can be passed through At.

In other embodiments, interior dielectric may include third layer dielectric material, and third layer dielectric material has contact The first surface of first layer dielectric material and the apparent surface for being covalently bound to hydrophobic layer.In such embodiments, first layer Dielectric material may include oxide, (or the other places this paper) as described above；Second layer dielectric material may include oxide or nitrogen Compound, (or described in elsewhere herein) as described above.In certain embodiments, third layer dielectric material may include oxidation Object, such as silica or other dielectric materials with siloxane group good combination.In certain embodiments, third layer dielectric Material passes through ALD deposition.In certain embodiments, third layer dielectric material has about 2nm to about 10nm or about 4nm to about 6nm Thickness.

Regardless of constituting the quantity of the layer of inner-dielectric-ayer, inner-dielectric-ayer can be with the total thickness of about 40nm to about 120nm Degree (for example, about 40nm to about 60nm, about 50nm to about 70nm, about 60nm to about 80nm, about 70nm to about 90nm, about 80nm extremely The range that about 100nm, about 90nm are limited to about 110nm, about 100nm to about 120nm or by the aforementioned endpoint of any two).It is similar Ground, dielectric layer can have about 50kOhm to about 150kOhm (for example, about 50kOhm to about 75kOhm, about 75kOhm are to about The model that 100kOhm, about 100kOhm are limited to about 125kOhm, about 125kOhm to about 150kOhm or by the aforementioned endpoint of any two It encloses).

In some embodiments, substrate may further include photoresponsive layer.Photoresponsive layer can have contact inner-dielectric-ayer The first side and contact the second side of at least one electrode.In certain embodiments, photoresponsive layer may include amorphous silicon hydride. In such embodiments, it can be reduced at light area with any region in the multiple regions of light beam irradiation light response layer The impedance of photoresponsive layer.In other embodiments, photoresponsive layer includes multiple conductors, and each conductor is via phototransistor switch It is controllably connected at least one electrode of substrate.

Include the embodiment of lid for wherein microfluidic device, the surface of the inwardly capsule of lid may include internal layer and It is covalently bound to the hydrophobic layer (that is, outer hydrophobic layer) of internal layer.Similar with the outer hydrophobic layer of substrate, the outer hydrophobic layer of lid can include It is covalently bound to the self-association molecule of internal layer, to form the hydrophobic monolayer of dense packing.Therefore, on outer hydrophobic layer may include Any self-association molecule of the outer hydrophobic layer for substrate of literary (or the other places this paper) description.In some embodiments, lid is outer Hydrophobic layer includes self-association molecule identical with the outer hydrophobic layer of substrate.In other embodiments, the outer hydrophobic layer of substrate has Outer hydrophobic layer of the self-association molecule of different type (or multiple types) as substrate.

In some embodiments, the outer hydrophobic layer of the inner surface of lid, which has, is less than 5 nanometers (for example, about 1.5 to 3.0 receive Rice) thickness.In some embodiments, the outer hydrophobic layer of the inner surface of lid can be patterned such that and outer hydrophobic layer Rest part is compared, and selection region is relative hydropathic.

In some embodiments, microfluidic device may include the capsule for having at least one microfluidic channel.In addition, envelope Shell may include at least one microfluidic chamber (or isolation fence) for being fluidly coupled to microfluidic channel.Limit microchannel and/ Or at least part of the substrate of chamber can have electrowetting configuration.Electrowetting configuration may be coupled to bias potential, and In such connection, change effective profit of any one of multiple corresponding regions of substrate surface (that is, drop actuation surfaces) Moisture performance.The wetting characteristics of substrate surface can fully be changed, so that liquid drop is on the surface of a substrate and in miniflow It is moved between body channel and chamber.

Holding area that in some embodiments, chamber (or isolation fence) may include being configured as keeping drop (for example, Isolated area) and by holding area be fluidly connected to microfluidic channel one (or more) bonding pad.First bonding pad can To be configured to allow for liquid drop to be moved between microfluidic channel and chamber.When there are the second bonding pad, it can be configured to Allow fluid to flow when liquid drop moves between microfluidic channel and holding area to discharge with pressure.In some embodiments In, capsule may further include the second microfluidic channel.In such embodiments, chamber may be coupled to the first microfluid Both channel and the second microfluidic channel.

In some embodiments, microfluidic channel can be with about 30 to about 200 microns or about 50 to about 150 microns of height Degree, wherein in normal direction in measurement height on the direction by the fluid flow direction in channel.In some embodiments, microfluid is logical Road has about 50 to about 1000 microns or about 100 to about 500 microns of width, wherein being flowed in by the fluid in channel in normal direction Width is measured on the direction in direction.

In some embodiments, chamber (or isolation fence) has the height substantially the same height with microfluidic channel. For example, chamber height can be about 30 to about 200 microns or about 50 to about 150 microns.In some embodiments, chamber (or protect Hold fence) section with about 100,000 to about 2,500,000 square microns or about 200,000 to about 2,000,000 square microns Area.In some embodiments, bonding pad (first, second etc.) is micro- with being opened from it with corresponding chamber and/or bonding pad The substantially the same height of the height of fluid channel.In some embodiments, bonding pad has about 50 to about 500 microns or about 100 to about 300 microns of width.

In some embodiments, microfluidic device may further include droplet generator.Droplet generator can by with Selectively drop by one or more of liquid mediums (for example, water-based liquid medium) is set to provide into capsule or capsule In interior microfluidic channel.Drop can include such as speck body, such as biological speck body (such as cell) or bead.It is alternative Ground or additionally, the drop may include reagent, such as lysis buffer, affinity reagent, detectable label, enzymatic mixture etc..

In some embodiments, microfluidic device includes being suitable for the culture chamber of the biological speck body of culture (for example, isolation is enclosed Column).Cultivating chamber can be located in capsule, and may be coupled to microfluidic channel.When culture chamber is located in capsule, envelope Shell may include perfusion microfluidic channel, which is configured as that fresh culture is made to flow through culture chamber so that new Waste in nutrients in fresh culture medium and culture chamber can swap (such as by nutrients into culture chamber The diffusion of diffusion and waste into culture medium).Perfusion channel can be separated with the microfluidic channel for being connected to droplet generator.

In some embodiments, Electrowetting device and electronic position-fixing device are integrated.For example, in some embodiments, miniflow Body equipment may include the substrate that there is electrowetting to configure, and a part for substrate may further include dielectrophoresis (DEP) Configuration.Therefore, substrate can be whole (monolithic) (monolithic).Optionally, microfluidic device or device can wrap It includes：With the first module of the first substrate or part, first substrate is configured with dielectrophoresis (DEP)；It is served as a contrast with second Second module at bottom or part, second substrate include electrowetting configuration.Such equipment may be considered that with binary (biplate) (duolithic) substrate, and may exist bridge between the first module or part and the second module or part, with It provides integrated with the associated function of each substrate and its specific configuration.The bridge may include the equipment for connecting two separation Pipeline or the like.Alternatively, the bridge may include making substrate close apposition (for example, in 2mm, 1.5mm, 1.0mm, 0.5mm or more It is small in) adhesive.In other alternative solution, bridge can be the nonfunctional area on integral type lining bottom, wherein nonfunctional area It is the place that substrate configuration is switched to another configuration (for example, DEP is configured) from a kind of configuration (such as electrowetting configuration).It is no matter micro- Fluid device is that with bulk substrate or have binary substrate (or even more body substrates), in electrowetting configuration and DEP configurations Each can be any this configuration known in the art or disclosed herein.For example, electrowetting configuration can be light Electrowetting (EWOD) configuration, unilateral electrowetting configuration etc. in electrowetting (OEW) configuration, dielectric.Similarly, DEP configurations can be with It is photoelectron tweezers (OET) configuration, such as by including amorphous silicon layer and/or photo-transistor arrays, being controlled by phototransistor Electrod-array, electric actuation the photoelectricity conductive substrate of electrod-array etc. provided.In some alternative embodiments, substrate can be with It is configured including electrowetting but lacks any additional configurations (for example, lacking dielectrophoresis (DEP) configuration).

Therefore, in some embodiments, single single unit system can combine the function of two devices.

On the other hand, the present invention provides the methods of the microfluidic device of the manufacture present invention.This method may include：By between It is attached to at least one electrode for being configured to connect to voltage source every element (for example, being made of microfluidic circuit material) Lid inner surface；The spacer element and lid are attached to the interior dielectric surface of substrate, the substrate has the company of being configured as It is connected at least one electrode of voltage source；And at least part and the lining by being vapor-deposited in the inner surface of the lid Hydrophobic layer is formed at least part of the interior dielectric surface at bottom.In certain embodiments, spacer element is clipped in the inner surface of lid Between the interior dielectric surface of substrate so that lid and substrate are essentially parallel from one to another orientation.Substrate, spacer element and Gai Ke are limited jointly Surely it is configured to keep the capsule of liquid medium.In certain embodiments, hydrophobic layer is deposited on the essentially all of the inner surface of lid The essentially all exposed region of the interior dielectric surface of exposed region and substrate is (that is, being facing inwardly toward the substantially complete of capsule On portion surface).In certain embodiments, hydrophobic layer is further deposited on the surface of inwardly capsule of spacer element.

In certain embodiments, hydrophobic layer includes forming certainly for the interior dielectric surface of the inner surface and substrate that are covalently bound to lid Molecule is closed, wherein self-association molecule forms the single layer of dense packing.In some embodiments, forming certainly by the deposition that is vapor-deposited Conjunction molecule includes each surface modified ligand and the link that surface modified ligand is directly or indirectly linked to inner-dielectric-ayer surface Group.Therefore, self-association molecule can be any self-association molecule above or described elsewhere herein.

On the other hand, the present invention is provided to handle the material of such as chemicals and/or biomaterial in microfluidic devices The method of material.In certain embodiments, the method includes：Fill the capsule or part of it of microfluidic device, the miniflow Body device includes substrate, lid and the spacer element that there is electrowetting to configure, they limit the envelope with the first liquid medium together Shell；Apply AC voltage potentials between at least one electrode of substrate and at least one electrode of lid；By the first of liquid medium Drop is introduced into the capsule, and the liquid medium of the drop is unmixing in first liquid medium；And by One drop applies electrowetting power and the first drop is moved to the desired locations in capsule.First liquid medium may include described herein Any first liquid medium, such as silicone oil, fluorinated oil or combinations thereof, and the first drop may include aqueous medium.

In some embodiments, the method may include first part's (such as microfluids by the first drop from capsule Channel) it is drawn in the second part (such as chamber) of capsule, vice versa.Aforementioned towing may include changing and the first drop Effective electro-wetting property in the region of contact and/or neighbouring substrate surface.It therefore, can be with the first liquid medium filling capsule Including filling microfluidic channel and chamber with the first liquid medium.

In some embodiments, microfluidic device includes droplet generator.This method may include using droplet generator Generate the first drop.In addition, the first drop can be introduced into capsule by droplet generator.The drop of generation can have about 100 skins Rise to the volume of 100 nanoliters or about 1 to 50 nanoliter.In some embodiments, the first drop may include such as bead or biology Speck body, cell secreta or the reagent of speck body (such as cell, vesica etc.).Bead can have all to interested material As cell secreta (such as antibody) or other biological molecule (such as nucleic acid, such as DNA, genomic DNA, mitochondrial DNA, RNA, mRNA, miRNA or any combination thereof) molecule with affinity.Drop may include the list of for example single biological cell A speck body or multiple speck bodies.For example, drop may include two to 20 or more speck bodies, such as bead.One In a little embodiments, drop may include reagent, such as cell lysis buffer solution, label (such as reagent of fluorescent marker), shine Reagent, enzymatic mixture etc..

In some embodiments, the method further include by second, third, the drops such as 4th be introduced into capsule and pass through by Electrowetting power be applied to drop and by second, third, the drops such as 4th be moved to desired locations in capsule.Second drop is removable It moves close to the position of the first drop, then combines drop with the first droplet coalescence to form first；Third drop can be moved into Close to the position of the first combination drop, then droplet coalescence is combined with first to form the second combination drop；4th drop can be with It is moved close to the position of the second combination drop, then droplet coalescence is combined with second to form third and combines drop；Etc..Often A additional drop may be embodied in unmixing in the first liquid medium but be situated between with the miscible fluid of the liquid medium of the first drop Matter.

In some embodiments, the first drop includes biological cell and the second drop includes reagent.Reagent can be thin Cellular lysate buffer solution dissolves biological cell in the first and second droplet coalescences.Alternatively, reagent can be fluorescent marker (example Such as, the antibody of fluorescent marker or other affinity reagents) or for luminescence assays reagent.Third drop may include reagent, such as There are one or more (for example, two to 20) of affinity to capture bead interested material.For example, interested Material can be antibody or nucleic acid, such as DNA, genomic DNA, mitochondrial DNA, RNA, mRNA, miRNA. Such capture pearl can optionally output be used for subsequent analysis from device.As second and third drop, the 4th liquid Drop can include reagent, be for example suitable for the enzymatic mixture of the reaction reacted such as reverse transcriptase reaction or whole genome amplification.

In some embodiments, the movement and merging of drop are related to using electrowetting power, including change connecing for substrate surface Effective electro-wetting property in the region of nearly drop, to mobile or merging drop.In certain embodiments, change substrate surface Effective electro-wetting property may include the activation electrowetting electrode at the region close to the substrate surface of drop.In some embodiments In, activation is related to for light pattern being directed to the region of substrate surface close to the electrowetting electrode at the region of the substrate surface of drop On.

From following attached drawing and detailed description, other aspects of the present invention and embodiment will be apparent.

Description of the drawings

Figure 1A shows general microfluidic device according to some embodiments of the present invention and with micro- for controlling and monitoring The system of the association control apparatus of fluid device.

Figure 1B is the vertical cross-section diagram of the microfluidic device with substrate, lid and spacer element, substrate, lid and spacer element The capsule for being configured as keeping the liquid drop of liquid medium and immiscible in liquid medium is formed together.Substrate, which has, to be permitted Perhaps the electrowetting configuration of drop is manipulated in capsule.

Fig. 1 C and Fig. 1 D show microfluidic device according to some embodiments of the invention.

Fig. 2A and 2B shows isolation fence according to some embodiments of the invention.

Fig. 2 C show detailed isolation fence according to some embodiments of the invention.

Fig. 2 D-2F show the isolation fence of some other embodiments according to the present invention.

Fig. 2 G show microfluidic device according to the ... of the embodiment of the present invention.

Fig. 2 H show the coating surface of microfluidic device according to the ... of the embodiment of the present invention.

Fig. 3 A show making together with microfluidic device and associated control device according to some embodiments of the invention The specific example of system.

Fig. 3 B show imaging device according to some embodiments of the invention.

Fig. 4 shows the example of the microfluidic device with EW configurations and DEP configurations with binary substrate.

Fig. 5 shows the example of the microfluidic device with EW configurations and DEP configurations with bulk substrate.

Fig. 6 is the horizontal sectional view of microfluidic device, may include that electrowetting as shown in Figure 1B configures, and it is wrapped The chamber and droplet generator for including multiple microfluidic channels, being opened from least one microfluidic channel.In this embodiment, one A microfluidic channel includes aqueous medium (shallower color), and the microfluidic channel for being connected to droplet generator includes non-aqueous Medium (deeper color).These chambers equally include aqueous medium or non-aqueous medium.

Fig. 7 is the horizontal sectional view of microfluidic device, may include that electrowetting as shown in Figure 1B configures, and it is wrapped The chamber and droplet generator for including multiple microfluidic channels, being opened from least one microfluidic channel.In this embodiment, one A microfluidic channel and first group of chamber include aqueous medium (shallower color), and the microfluid for being connected to droplet generator is logical Road and second group of chamber include hydrophobic medium (dark color).Fig. 6 shows the modification of embodiment illustrated in fig. 5, wherein wrapping Each chamber of aqueous medium is located immediately at the channel of the hydrophobic medium with the corresponding chambers for carrying out self-contained hydrophobic medium On.

Fig. 8 is the figure for the method for handling the biological speck body in microfluidic device.

Fig. 9 is the method for the substrate that can be applied to manufacture microfluidic device, and microfluidic device, which has, to be configured with electrowetting First part and the second part configured with dielectrophoresis.

Figure 10-18 provides the vertical cross-section diagram of the substrate according to the method processing described in Fig. 9.

Figure 19 A are regarding for what the electrical addressing operation in terms of function indicated according to the embodiment combined shown in Figure 17 Figure.

Figure 19 B are regarding for what the electrical addressing operation in terms of function indicated according to the embodiment combined shown in Figure 17 Figure.

Figure 20 A-20C are the photographs of the movement of the aqueous drop according to an embodiment of the invention on modified microfluidic surface Piece indicates.

Specific implementation mode

Present specification describes exemplary embodiment of the present invention and application.However, that the present invention is not restricted to these is exemplary The mode that embodiment and application or exemplary embodiment and application are operated or described herein.In addition, attached drawing can show letter Change or partial view, and the size of the element in attached drawing may be amplified or otherwise disproportionate.In addition, by In herein use term " ... on ", " being attached to ", " being connected to ", " being coupled to " or similar word, an element is (for example, material Material, layer, substrate etc.) can " " another element "upper", " being attached to ", " being connected to " or " being coupled to " another element, no matter one A element whether directly on another element, be attached to, be connected or coupled to another element, or in an element and another There are one or more intermediary elements between a element.In addition, unless the context indicates otherwise, otherwise direction (for example, top, Lower section, top, bottom, side, upper and lower, downside, upside, top, lower part, level, vertical, " x ", " y ", " z " etc.) (if If) be opposite and only provide by way of example, and limited for convenience of description with discussing.This Outside, when referring to element list (for example, element a, b, c), such reference is intended to include any one of the element listed The element individually listed, all or fewer than the combination of any combinations for the element listed and/or all elements listed.Explanation Chapter construction in book is not intended to limit any factor combination of discussion only convenient for checking.

As used herein, " substantially " mean to be sufficient to expected purpose.Therefore, term " substantially " allows absolute or complete Small, the inapparent variation of beautiful state, size, measurement, result etc., such as desired by those of ordinary skill in the art , but not significantly affect overall performance.Come in use, " base when about numerical value or parameter or the feature that can be indicated with numerical value In sheet " mean in 10.

Term " multiple " means more than one.

As it is used herein, term " multiple " can be 2,3,4,5,6,7,8,9,10 or more.

As it is used herein, term " setting " covers the meaning of its " being located at ".

As used herein, " microfluidic device " or " microfluidic device " be include one or more discrete microfluidic circuits Equipment, the discrete microfluidic circuit be configured as accommodate fluid, each microfluidic circuit include fluid interconnection loop element (including but not limited to region, flow region, channel, chamber and/or fence) and (for the microfluidic device including lid) is extremely Few two ports, at least two port are configured to allow for fluid (the speck body and optionally, to suspend in a fluid) to flow Enter and/or flow out microfluidic device.Typically, the microfluidic circuit of microfluidic device will include at least one microfluidic channel and At least one chamber, and the fluid that about 1mL volumes will be contained less than, for example, less than about 750,500,250,200,150,100, 75,50,25,20,15,10,9,8,7,6,5,4,3 or 2 μ L.In certain embodiments, microfluidic circuit accommodate about 1-2,1-3, 1-4、1-5、2-5、2-8、2-10、2-12、2-15、2-20、5-20、5-30、5-40、5-50、10-50、10-75、10-100、 20-100,20-150,20-200,50-200,50-250 or 50-300 μ L.

As used herein, " receive fluid device " or " receive fluid means " is a kind of microfluidic device, is had comprising at least The microfluidic circuit of one loop element, the loop element are configured as accommodating the fluid of less than about 1 μ L volumes, are, for example, less than About 750,500,250,200,150,100,75,50,25,20,15,10,9,8,7,6,5,4,3,2,1nL or less.Receive fluid Equipment may include multiple loop elements (for example, at least 2,3,4,5,6,7,8,9,10,15,20,25,50,75,100,150, 200、250、300、400、500、600、700、800、900、1000、1500、2000、2500、3000、3500、4000、4500、 5000,6000,7000,8000,9000,10,000 or more).In certain embodiments, one at least one loop element A or more (such as all) be configured as accommodating about 100pL to 1nL, 100pL to 2nL, 100pL to 5nL, 250pL extremely 2nL, 250pL to 5nL, 250pL to 10nL, 500pL to 5nL, 500pL to 10nL, 500pL to 15nL, 750pL to 10nL, Fluids of the 750pL to 15nL, 750pL to 20nL, 1 to 10nL, 1 to 15nL, 1 to 20nL, 1 to 25nL or 1 to 50nL volumes. In other embodiment, one or more (such as whole) at least one loop element are configured as accommodating about 20nL extremely 200nL, 100 to 200nL, 100 to 300nL, 100 to 400nL, 100 to 500nL, 200 to 300nL, 200 to 400nL, 200 to 500nL, 200 to 600nL, 200 to 700nL, 250 to 400nL, 250 to 500nL, 250 to 600nL or 250 are to 750nL volumes Fluid.

" microfluidic channel " or " flow channel " refers to having to be considerably longer than horizontal and vertical size as used herein The flow region of the microfluidic device of length.For example, flow channel can be at least 5 times of length of horizontally or vertically size, example Such as at least 10 times of length, at least 25 times of length, at least 100 times of length, at least 200 times of length, at least 500 times of length, at least 1, 000 times of length, at least 5,000 times of length or longer.In some embodiments, the length of flow channel is from about 50,000 microns Any range in the range of about 500,000 microns, including therebetween.In some embodiments, horizontal size is at about 100 microns To in the range of about 1000 microns (for example, about 150 to about 500 microns), and vertical dimension is at about 25 microns to about 200 microns In the range of, for example, from about 40 to about 150 microns.Notice that flow channel there can be various differences in microfluidic device Space configuration, therefore be not limited to perfect linear element.For example, flow channel may include have it is any in following configuration One one or more part：Curve, bending, spiral, inclination, decline, fork branch (for example, multiple and different flow paths) And any combination of them.In addition, flow channel there can be different sectional areas along its path, widens and shrink with wherein Desired fluid flowing is provided.

As it is used herein, term " barrier " is commonly referred to as the structure of sufficiently large bulge or similar type, Two shifting different zones or loop element between of the target speck body in microfluidic device is prevented with part (but incomplete) It is dynamic.Two different zones/loop elements can be that such as microfluid isolation fence and microfluidic channel or microfluid isolation are enclosed The bonding pad on column and isolated area.

As it is used herein, term " contraction " typically refers to loop element (or two circuit members in microfluidic device Interface between part) width narrow.Contraction flow region can for example be located at the boundary between microfluid isolation fence and microfluidic channel Place, or the intersection between the isolated area and bonding pad that microfluid completely cuts off fence.

As it is used herein, term " transparent " refer to allow visible light by by substantially will not light by when change The material of light.

As it is used herein, term " speck body " typically refers to any speck that can be detached and collect according to the present invention Body.The non-limiting example of speck body includes：Abiotic speck body, such as particle, microballon (for example, polystyrene bead, Luminex^TMBead etc.), magnetic bead, micro- stick, microfilament, quantum dot etc.；Biological speck body, such as cell (such as embryo, ovum mother are carefully Born of the same parents, ovum, spermatid, the cell from tissue separation, eukaryocyte, blastema, zooblast, mammalian cell, people are thin Born of the same parents, immunocyte, hybridoma, the cell of culture, the cell from cell line, cancer cell, infection cell, transfection and/or conversion Cell, report cell, prokaryotic cell etc.)；Biological cell device；Vesica or compound；Synthesize vesica；Liposome (such as synthesize Or be derived from film preparation)；Lipid nanometer bar (such as Ritchie (2009) " Reconstitution of Membrane Proteins in Phospholipid Bilayer Nanodiscs (rebuilding memebrane protein in phospholipid bilayer nanometer disk) ", Methods Enzymol., 464：Described in 211-231) etc.；Or without life speck body and biological speck body (for example, attachment In the coated microballon of microballon, liposome of cell, the coated magnetic bead of liposome etc.) combination.Bead can further have altogether Other parts/molecule of valence or non-covalent linking, such as fluorescent marker, protein, small molecule signal section, antigen can be used In the chemical/biological substance of measurement.

As used herein, term " maintain (one or more) cell " refer to provide comprising fluid and gas component and The environment on optional surface provides the condition kept needed for cell survival and/or amplification.

" component " of fluid media (medium) is any chemistry or biochemical molecular present in medium, including solvent molecule, ion, small Molecule, antibiotic, nucleotide and nucleosides, nucleic acid, amino acid, peptide, protein, sugar, carbohydrate, lipid, aliphatic acid, courage are solid Alcohol, metabolin etc..

As herein with respect to used in fluid media (medium), " diffusion (diffuse) " and " diffusion (diffusion) " refers to stream The component of body medium along concentration gradient thermodynamic motion.

Term " flowing of medium " refer to mainly due to diffusion other than any mechanism caused by fluid media (medium) a large amount of shiftings It is dynamic.For example, due to the pressure difference between point, the flowing of medium may relate to fluid media (medium) and be moved to another point from a bit.It is this Flowing may include the continuous of liquid, pulse, periodicity, random, interval or reciprocal flowing, or any combination thereof.When a kind of stream When body medium flows into one other fluid medium, turbulent flow and the mixing of medium may be caused.

Phrase " not flowing substantially " refers to that the flow rate of fluid media (medium) averagely obtained at any time is less than material (example Such as, analytes of interest analytes) diffusion of components to fluid media (medium) in or fluid media (medium) in rate.The diffusion of the component of this material Rate can depend on the intensity that interacts between the size and component and fluid media (medium) of such as temperature, component.

As herein in regard to used in the different zones in microfluidic device, phrase " fluidly connecting " refers to, when not same district When domain is substantially saturated with fluid (such as fluid media (medium)), fluidly connecting to form a single fluid in each region.This The fluid (or fluid media (medium)) being not meant in different zones must be the same in composition.But the difference of microfluidic device Fluid in fluid junction can have different compositions (such as the solute of various concentration, such as protein, carbon hydrate Object, ion or other molecules), it is moved down along its respective concentration gradient as solute in flowing and/or fluid flows through this Equipment.

Microfluid (or receive fluid) equipment may include " being swept " region and " not being swept " region.Such as this paper institutes It uses, " being swept " region includes the loop element of one or more fluid interconnections of microfluidic circuit, works as fluid stream When crossing microfluidic circuit, each loop element undergoes medium flow field.The loop element in the region being swept may include such as region, Channel and all or part of chamber.As it is used herein, " not being swept " region include one of microfluidic circuit or The loop element of more fluid interconnections, when fluid flows through microfluidic circuit, each loop element does not suffer from fluid substantially Flux.The region not being swept can be fluidly connected to the region being swept, if fluidly connect be configured to diffusion but It there is no that medium flows between the region being swept and the region not being swept.Therefore microfluidic device can be configured to Substantially not inswept region and the medium flow field in the region that is swept are kept apart, while not being swept in the region being swept and Region between substantially only allow divergent fluid be connected to.For example, the flow channel of microfluidic device is the region being swept Example, and the isolated area (being detailed further below) of microfluidic device is the example in the region not being swept.

As it is used herein, " flow region " refers to one or more loop elements fluidly connected (such as one Or multiple channels, one or more regions, one or more chambers etc.), it limits media flow and is subjected to media flow track. Therefore flow region is the example in the region of microfluidic device being swept.Other loop elements are (for example, the area not being swept Domain) flowing of medium that can fluidly connect with the loop element including flow region, and be subjected in flow region.

As used herein, " alkyl " refers to the linear chain or branched chain hydrocarbon chain radial configuration being only made of carbon and hydrogen atom, no Containing unsaturation, there is 1-6 carbon atom (such as C1-C6 alkyl).No matter when appear in herein, the digital model of such as " 1 to 6 " It refers to each integer in given range to enclose；Such as " 1 to 6 carbon atom " refers to that alkyl can be by 1 carbon atom, 2 carbon originals The compositions such as son, 3 carbon atoms, until and include 6 carbon atoms, although this definition is also contemplated by term " alkyl " and does not specify numerical value model The case where enclosing.In some embodiments, it is C1-C3 alkyl.Typical alkyl include but is not limited to methyl, ethyl, propyl, Isopropyl, normal-butyl, isobutyl group, sec-butyl isobutyl group, tertiary butyl, amyl, isopentyl, neopentyl, hexyl etc..Alkyl passes through list It is keyed to the rest part of molecule, for example, methyl (Me), ethyl (Et), n-propyl, 1- Methylethyls (isopropyl), positive fourth Base, n-pentyl, 1,1- dimethyl ethyls (tertiary butyl), hexyl etc..

Unless being otherwise noted in the description, otherwise alkyl can optionally be replaced by one or more substituent groups, institute Substituent group is stated independently to be：Aryl, aryl alkyl, heteroaryl, heteroaryl alkyl, hydroxyl, halogen, cyano, trifluoromethyl, trifluoro Methoxyl group, nitro, trimethylsilyl ,-OR ' ,-SR ' ,-OC (O)-R ' ,-N (R ') 2 ,-C (O) R ' ,-C (O) OR’、—OC(O)N(R’)2、—C(O)N(R’)2、—N(R’)C(O)OR’、—N(R’)C(O)R’、—N(R’)C(O)N(R’) 2, N (R ') C (NR ') N (R ') 2 ,-N (R ') S (O) tR ' (wherein t is 1 or 2) ,-S (O) tOR ' (wherein t is 1 or 2) ,-S (O) tN (R ') 2 (wherein t is 1 or 2) or PO3 (R ') 2, wherein each R' independently is hydrogen, alkyl, fluoro-alkyl, aryl, virtue Alkyl, Heterocyclylalkyl or heteroaryl.

As mentioned in this article, fluorinated alkyl part is that one or more hydrogen of moieties are replaced by fluoro substituents Moieties.There are perfluoroalkyl moieties the whole hydrogen for being connected to moieties to be replaced by fluoro substituents.

As mentioned in this article, " halogenated " is partly bromine, chlorine or fluorine part.

As mentioned in this article, " olefinic " compound is the organic molecule containing " alkene " part.Hydrocarbon fraction refer to by The group of at least two carbon atoms and at least one carbon-to-carbon double bond composition.The non-olefinic part of molecule can be any classification Organic molecule, and in some embodiments, may include alkyl or fluorination (including but not limited to fluoridized) moieties, Any of which one can be further substituted.

As used herein, " hydrophobic monolayer of dense accumulation " refers to single layer of hydrophobic molecule, is sufficiently closely packed together To resist the insertion and/or intrusion of polar molecule (such as water, ion and other charge species).

As used herein：" μm " (or " um ") refers to micron；“μm³" refer to cu μ m；" pL " refers to picoliters, and " nL ", which refers to, to be received It rises；And " μ L " (or " uL ") refers to microlitre.

Loading method.Speck body (such as biological speck body and/or bead) is loaded into the different zones of microfluidic device In can be related to using fluid flowing, gravity, dielectrophoresis (DEP) power, electrowetting power, magnetic force, such as institute here Description.Can be such as configured by photoelectricity tweezers (OET) optically to generate DEP power, and/or such as by activationary time/ Electrode/electro polar region domain in spatial framework and electric power generates DEP power.Similarly, photoelectricity wetting (OEW) configuration can such as be passed through Optically to generate electrowetting power, and/or such as electric power production by electrode/electro polar region domain in activationary time/spatial framework Raw electrowetting power.

Microfluidic device for operating and observing these equipment and system.Figure 1A shows that can be used for controlling microfluid sets For 100 and the general sample of the microfluidic device 100 and system 150 of the movement of speck body therein and/or drop.It shows micro- The perspective view of fluid device 100, the partial cut with its lid 110 is to provide the partial view of microfluidic device 100.Miniflow Body equipment 100 generally includes microfluidic circuit 120, which includes fluid mass 106, and fluid media (medium) 180 can lead to The fluid mass 106 flowing is crossed, optionally one or more speck body (not shown) are carried in microfluidic circuit 120 And/or pass through microfluidic circuit 120.Although showing single microfluidic circuit 120 in figure 1A, suitable microfluid is set Standby may include microfluidic circuit as multiple (such as 2 or 3).Anyway, microfluidic device 100 can be configured to Nanofluidic devices.In embodiment shown in figure 1A, microfluidic circuit 120 include multiple microfluids isolation fence 124,126, 128 and 130, each microfluid isolation fence has the single opening being in fluid communication with flow region 106.As begged for further below Opinion, it includes various feature and structure that microfluid, which completely cuts off fence, these feature and structure have been optimized for protecting speck body It holds in such as microfluidic device of microfluidic device 100, even if when medium 180 flows through flow region 106.However, turning To before aforementioned, the brief description of microfluidic device 100 and system 150 is provided.

As shown generally in Figure 1A, microfluidic circuit 120 is limited by capsule 102.Although capsule 102 can with physique at Different configurations, but in example shown in figure 1A, capsule 102 is depicted as including support construction 104 (such as base portion), miniflow Body loop structure 108 and lid 110.However, in certain embodiments, capsule 102 can lack lid 110, and microfluidic circuit 120 can be limited by support construction 104 and microfluidic circuit structure 108.Support construction 104,108 and of microfluidic circuit structure (optionally) lid 110 can be attached to each other.For example, the inner surface in support construction 104 can be arranged in microfluidic circuit structure 108 On 109, and lid 110 can be arranged above microfluidic circuit structure 108.With support construction 104 and (optionally) lid 110 1 It rises, microfluidic circuit structure 108 can limit the element of microfluidic circuit 120.

As shown in Figure 1A, support construction 104 can be located at 120 bottom of microfluidic circuit, and lid 110 can be located at miniflow The top in body circuit 120.Alternatively, support construction 104 and lid 110 can be with other azimuth configurations.For example, support construction 104 can With positioned at the top of microfluidic circuit 120, and lid 110 can be located at the bottom of microfluidic circuit 120.It is anyway possible to There are one or more port 107, each port include the channel for entering or leaving capsule 102.The example in channel include valve, Gate, through-hole etc..As shown, port 107 is the through-hole formed by the gap in microfluidic circuit structure 108.However, port 107 can be located in the other component of capsule 102, such as lid 110.A port 107, but microfluidic circuit are only shown in Figure 1A 120 can have two or more ports 107.For example, there may be used as that fluid enters the entrance of microfluidic circuit 120 the Single port 107, and there may be the second ports 107 for the outlet that microfluidic circuit 120 is left as fluid.Make port 107 The direction that fluid flows move region 106 can be depended on by still being exported for entrance.

Support construction 104 may include one or more electrode (not shown) and substrate or the substrate of multiple interconnection.Lining Bottom can be any suitable substrate known in the art.For example, support construction 104 may include one or more semiconductors Substrate, each semiconductor substrate are electrically connected at least one of one or more electrodes (for example, all or part of semiconductor Substrate may be electrically connected to single electrode).Alternatively, support construction 104 may include including the printing of one or more electrodes Circuit board assemblies (" PCBA ").In other embodiments, support construction 104 may include be mounted on PCBA on substrate (for example, Semiconductor substrate).

Microfluidic circuit structure 108 can limit the loop element of microfluidic circuit 120.Such loop element can wrap It includes when microfluidic circuit 120 is full of fluid, space or region that can be to be interconnected on fluid, such as (it can be wrapped flow region Include or can be one or more flow channels), chamber, fence, trap etc..In microfluidic circuit 120 shown in figure 1A In, microfluidic circuit structure 108 includes frame 114 and microfluidic circuit material 116.Frame 114 can be sealed partially or completely Close microfluidic circuit material 116.Frame 114 can be the relative stiffness for example essentially around microfluidic circuit material 116 Structure.For example, frame 114 may include metal material.Alternatively, microfluidic circuit structure 108 can lack frame.For example, micro- Fluid circuit structure 108 can be made of microfluidic circuit material 116 or is substantially made of microfluidic circuit material 116.

Can with cavity etc. come pattern microfluidic circuit material 116 with limit microfluidic circuit 120 loop element and Interconnection.Microfluidic circuit material 116 may include flexible material, such as flexible polymer (such as rubber, plastics, elastomer, silicon Resin, dimethyl silicone polymer (" PDMS ") etc.), can be ventilative.It may make up the material of microfluidic circuit material 116 Other examples include moulding etchable material (such as photo-patterned polysiloxanes or " PPS "), the light of glass, such as silicones Cause resist (such as SU8) etc..In some embodiments, such material (and therefore microfluidic circuit material 116) can be with It is rigid and/or substantially air impermeable.Anyway, microfluidic circuit material 116 can be arranged in support construction 104 And (optionally) it is arranged inside frame 114.

Lid 110 can be the integral part of microfluidic circuit material 116 and/or frame 114.Alternatively, lid 110 can be knot Different element on structure, as shown in Figure 1A.Lid 110 may include it is identical as frame 114 and/or microfluidic circuit material 116 or Different materials.Similarly, as shown, support construction 104 can be separated with microfluidic circuit material 116 or frame 114 Structure or microfluidic circuit material 116 or frame 114 component part.Similarly, 116 He of microfluidic circuit material Frame 114 (if present) can be separated structure as shown in Figure 1A or mutually isostructural integral part.

In some embodiments, lid 110 may include rigid material.Rigid material can be glass or have similar quality Material.In some embodiments, lid 110 may include deformable material.Deformable material can be polymer, such as PDMS.In some embodiments, lid 110 may include rigidity and deformable material.For example, one or more portions of lid 110 It may include rigid with lid 110 to divide (for example, one or more parts positioned at 124,126,128,130 top of isolation fence) The deformable material that property material connects.In some embodiments, lid 110 may further include one or more electrodes.It should One or more electrodes may include conductive oxide, such as tin indium oxide (ITO), can be applied to glass or similar Insulating materials on.Alternatively, one or more electrode can be the flexible electrode in embedded deformable material, such as singly Wall nanotube, many walls nanotube, nano wire, conductive nano-particles cluster or combinations thereof, the deformable material such as polymer (example Such as, PDMS).For example, the flexible electrode that can be used for microfluidic device is described in US2012/0325665 (Chiou et al.), Its content is incorporated herein by reference.In some embodiments, lid 110 can be by modification (for example, by coating or adjusting inwardly The all or part on the surface towards microfluidic circuit 120) with support drop mobile and/or cell adherence, cell viability and/or Cell growth.The modification may include the coating of synthesis or natural polymer or have covalently bound molecule (for example, self-association Molecule) conditioning surface.In some embodiments, lid 110 and/or support construction 104 can be transparent to light.Lid 110 It can also include the material (for example, PDMS or PPS) of at least one gas-permeable.

Figure 1A also shows the system 150 for operating and controlling microfluidic device (such as microfluidic device 100).System 150 include power supply 192, imaging device 194 (part for being not shown but can be image-forming module 164) and reclining device 190 (be not shown, but can be a part for tilt module 166).

Power supply 192 can provide electric power to microfluidic device 100 and/or reclining device 190, provide biased electrical as needed Pressure or electric current.For example, power supply 192 may include one or more exchanges (AC) and/or direct current (DC) voltage or current source.At As equipment 194 may include the equipment for capturing the image in microfluidic circuit 120, such as digital camera.In some cases Under, imaging device 194 further includes the detector (for example, being used for low illumination applications) for having quick frame rate and/or high sensitivity. Imaging device 194 can also include for stimulating radiation and/or light beam to be directed in microfluidic circuit 120 and collected from micro- Fluid circuit 120 (or in which comprising speck body) reflection or the radiation of transmitting and/or the mechanism of light beam.The light beam of transmitting can be with It in the visible spectrum, and can be for example including fluorescent emission.The reflected beams may include from LED or wide spectrum lamp (such as Mercury lamp (such as high-pressure sodium lamp) or xenon arc lamp) reflection transmitting.As discussed about Fig. 3 B, imaging device 194 can be into One step includes microscope (or optical train), may include or including eyepiece.

System 150 further comprises that reclining device 190, reclining device 190 are configured to surround one or more rotations Axis rotates microfluidic device 100.In some embodiments, reclining device 190 be configured around the support of at least one axis and/ Or keep include microfluidic circuit 120 capsule 102 so that microfluidic device 100 (and therefore microfluidic circuit 120) can be with It is maintained at level orientation (i.e. relative to x-axis and y-axis at 0 °), vertical orientations (i.e. in 90 ° relative to x-axis and/or y-axis) or therebetween Any orientation.Microfluidic device 100 (and microfluidic circuit 120) is referred to herein as microfluid relative to the orientation of axis " inclination " of equipment 100 (and microfluidic circuit 120).For example, reclining device 190 can make microfluidic device 100 relative to x-axis Or y-axis tilt 0.1 °, 0.2 °, 0.3 °, 0.4 °, 0.5 °, 0.6 °, 0.7 °, 0.8 °, 0.9 °, 1 °, 2 °, 3 °, 4 °, 5 °, 10 °, 15 °, 20 °, 25 °, 30 °, 35 °, 40 °, 45 °, 50 °, 55 °, 60 °, 65 °, 70 °, 75 °, 80 °, 90 ° or any angle therebetween.It is horizontal Orientation (and x-axis and y-axis) is defined as perpendicular to the vertical axis defined by gravity.Reclining device can also make microfluidic device 100 (and microfluidic circuits 120) relative to x-axis and/or y-axis tilt it is any be more than 90 ° of angle, or make microfluidic device 100 (and microfluidic circuit 120) tilts 180 ° relative to x-axis or y-axis, to invert (and the microfluidic circuit of microfluidic device 100 completely 120).Similarly, in some embodiments, reclining device 190 make microfluidic device 100 (and microfluidic circuit 120) surround by The rotation axis that 106/ channel 122 of flow region or some other parts of microfluidic circuit 120 limit tilts.

In some cases, microfluidic device 100 is inclined to vertical orientations so that 106/ channel 122 of flow region is located at Above or below one or more isolation fences.Term " top " as used herein indicates 106/ channel of flow region 122 are positioned higher than one or more isolation fences (that is, being located at flow region 106/ on the vertical axis limited by gravity Object in the isolation fence of the top of channel 122 will be with gravitional force more higher than object in flow region/channel).This The term " lower section " that text uses indicates that 106/ channel 122 of flow region is located below one on the vertical axis limited by gravity A or more isolation fence will be (that is, the object in the isolation fence of 106/ channel of flow region, 122 lower section will have than stream The lower gravitional force of object in dynamic region/channel).

In some cases, reclining device 190 makes microfluidic device 100 around being parallel to 106/ channel 122 of flow region Axis tilt.In addition, microfluidic device 100 can tilt to the angle less than 90 ° so that 106/ channel 122 of flow region Above or below one or more isolation fences, without positioned at the surface or underface of isolation fence.At it In the case of him, reclining device 190 makes microfluidic device 100 surround the axis inclination perpendicular to 106/ channel 122 of flow region. In the case of other, reclining device 190 makes microfluidic device 100 around both not parallel or be not orthogonal to 106/ channel of flow region 122 axis tilts.

System 150 can also include medium source 178.Medium source 178 (such as container, reservoir etc.) may include multiple portions Divide or container, each part or container are used to accommodate different fluid media (medium)s 180.Therefore, as shown in Figure 1A, medium source 178 can To be to be located at the equipment detached except microfluidic device 100 and with microfluidic device 100.Alternatively, medium source 178 can whole or portion Divide in the capsule 102 of microfluidic device 100.For example, medium source 178 may include one as microfluidic device 100 The reservoir divided.

The control that Figure 1A also shows a part for composition system 150 and can be used in combination with microfluidic device 100 Describe with the exemplary simplified block diagram of monitoring device 152.As shown, the example of this control and monitoring device 152 includes master Controller 154, the medium module 160 for controlling medium source 178, for control speck body in microfluidic circuit 120 and/or The power plant module 162 of movement and/or the selection of medium (for example, drop of medium), for control for capture images (such as count Word image) imaging device 194 (for example, camera, microscope, light source or any combination thereof) image-forming module 164, Yi Jiyong In the tilt module 166 of control reclining device 190.Control device 152 can also include for controlling, monitoring or execute about micro- Other modules 168 of other functions of fluid device 100.As shown, equipment 152 can operationally with display equipment 170 (or further comprising them) is coupled with input-output apparatus 172.

Master controller 154 may include control module 156 and digital storage 158.Control module 156 may include for example Digital processing unit, the digital processing unit are configured as the machine according to the non-transitory data or signal being stored as in memory 158 Device executable instruction (for example, software, firmware, source code etc.) is operated.Alternatively or additionally, control module 156 May include hard-wired digital circuit and/or analog circuit.Medium module 160, image-forming module 164, tilts power plant module 162 Module 166 and/or other modules 168 can be configured similarly.Therefore, can by as above configure master controller 154, be situated between Any one of matter module 160, power plant module 162, image-forming module 164, tilt module 166 and/or other modules 168 or more It is multiple to execute function, the process discussed in this article for performed by microfluidic device 100 or any other microfluidic device Action, action or process steps.Similarly, master controller 154, medium module 160, power plant module 162, image-forming module 164, incline Inclined module 166 and/or other modules 168 can be communicatively coupled, with send and receive any function being discussed herein, process, Data used in action, action or step.

Medium module 160 controls medium source 178.For example, medium module 160 can control medium source 178 will select Fluid media (medium) 180 is input in capsule 102 (for example, by ingress port 107).Medium module 160 can also be controlled from capsule Medium (for example, passing through output port (not shown)) is removed in 102.Therefore one or more media can be selectively input It is removed in microfluidic circuit 120 or from microfluidic circuit 120.Medium module 160 can also control in microfluidic circuit 120 The flowing of fluid media (medium) 180 in 106/ channel 122 of flow region.For example, in some embodiments, medium module 160 is being incited somebody to action Speck body or bead are loaded into isolation fence (using gravity, electrowetting (EW) power, dielectrophoresis (DEP) power or combinations thereof) it The preceding flowing for stopping medium 180 in 106/ channel 122 of flow region and across capsule 102.

Power plant module 162 can be configured as the choosing of speck body and/or medium drop in control microfluidic circuit 120 It selects, trap and moves.As discussed in detail, capsule 102 may include electrowetting (EW) configuration, and such as photoelectricity soaks (OEW) configuration, electrowetting (EWOD) configuration on dielectric, the configuration of single side electrowetting etc..In certain embodiments, capsule 102 It may further include dielectrophoresis (DEP) configuration, photoelectron tweezers (OET) configuration, electrically driven (operated) DEP configurations etc..Power Module 162 can be controlled by this EW and/or DEP configurations included electrode and/or transistor (for example, phototransistor) Activation, to select and move 106/ channel 122 of flow region and/or completely cut off the medium in fence 124,126,128,130 Speck body and/or drop.

Image-forming module 164 may be controlled to as 194 (not shown) of equipment.For example, image-forming module 164 can receive and process Image data from imaging device 194.Image data from imaging device 194 may include being captured by imaging device 194 Any kind of information (for example, the existence or non-existence of speck body, the drop of medium, label (fluorescence labels etc.) Accumulation).Using the information captured by imaging device 194, image-forming module 164 can further calculate object (for example, speck body, Medium drop etc.) movement rate in microfluidic device 100 of position and/or these objects.

Tilt module 166 can control the banking motion of 190 (not shown) of reclining device.In addition, tilt module 166 can be with Ramp rates and timing are controlled, such as speck body is transferred to by one or more isolation fences by gravity with optimization.It tilts Module 166 is communicatively coupled with image-forming module 164 and describes the speck body in microfluidic circuit 120 and/or medium drop to receive Movement data.Using this data, tilt module 166 can adjust the inclination of microfluidic circuit 120, micro- so as to adjustment Object and/or dielectric fluid drop in the rate moved in microfluidic circuit 120.Tilt module 166 can also be changed using this data The position of speck body and/or medium drop in generation ground adjustment microfluidic circuit 120.

In example shown in figure 1A, microfluidic circuit 120 is illustrated as including substantially being made of microfluidic channel 122 Single flow region 106.Each in isolation fence 124,126,128 and 130 includes leading to 106/ channel of flow region 122 single opening, is surrounded in addition to this so that fence can be by speck body and 106/ channel of flow region in fence 180 substantial barrier of speck body and/or fluid media (medium) in 122 or in other fences.The wall for completely cutting off fence can be from base portion Inner surface 109 extends to the inner surface of lid 110, to be conducive to this isolation.Fence is opened to 106/ channel 122 of flow region Mouth can be relative to the angled orientation of flowing of the fluid media (medium) 180 in 106/ channel 122 of flow region so that fluid media (medium) 180 flowing is not directed into fence.The flowing can be for example tangent or orthogonal with the plane of the opening of fence.In some feelings Under condition, fence 124,126,128 and/or 130 is configured as physically surrounding one or more micro- in microfluidic circuit 120 Object.Isolation fence according to the present invention may include in order to EW, OEW, DEP and/or OET power, fluid flowing and/or gravity Be used together and the variously-shaped, surface optimized and feature, as will be discussed in detail below.

Microfluidic circuit 120 may include any amount of microfluid isolation fence.Although showing five isolation fences, But microfluidic circuit 120 can have fewer or more isolation fences.As shown, the microfluid of microfluidic circuit 120 every Exhausted fence 124,126,128 and 130 includes respectively different feature and shape, these features and shape can be microfluidic device 100 provide the manipulation to the drop of speck body and/or fluid media (medium) useful one or more benefits.Therefore, in some realities It applies in example, microfluidic circuit 120 may include multiple microfluid isolation fences, and two of which or more isolation fence includes carrying For the different structure and/or feature of different benefits.However, in some embodiments, microfluidic circuit 120 includes multiple identical Microfluid completely cuts off fence.Can be used for manipulating speck body and/or the microfluidic device of medium drop may include any isolation fence 124,126,128 and 130 or their modification, including it those of is configured to as shown in Fig. 2 B, 2C, 2D, 2E and 2F fence, it is as follows It is described.

In embodiment shown in figure 1A, single flow region 106 is shown.However, other realities of microfluidic device 100 It can includes multiple flow regions 106 to apply example, and each flow region 106 be configured to fluid and flow through microfluidic device 100 to carry For the path of separation.Microfluidic circuit 120 includes the inlet valve being in fluid communication with flow region 106 or port 107, thus fluid Medium 180 can enter 106/ channel 122 of flow region via ingress port 107.In some cases, flow region 106 wraps Include single flow path.In other cases, flow region 106 includes multiple flow paths (such as 2,3,4,5,6 or more), Each may include microchannel (for example, as channel 122).In multiple flow paths two or more (such as It can all) be substantially parallel to each other.For example, flow region 106 is segmented into multiple parallel channel (such as channels 122).In certain embodiments, flow region 106 (and one or more channels for including by flow region) is with zigzag Pattern arranges that thus flow region 106 passes through microfluidic device 100 two or more times on alternating direction.In some feelings Under condition, the fluid media (medium) in each flow region 106 is flowed along at least one of direction direction forward or backwards.In some feelings Under condition, multiple isolation fences (for example, relative to 106/ channel 122 of flow region) are configured so that isolation fence can be with target Speck body parallel boot.

In some embodiments, microfluidic circuit 120 further includes one or more speck body traps 132.Trap 132 are generally formed in the wall on the boundary for constituting 106/ channel 122 of flow region, and can be with one or more microfluids Completely cut off the opening relative positioning of fence 124,126,128 and 130.In some embodiments, trap 132 is configured as from flowing 106/ channel 122 of region receives or captures single speck body.In some embodiments, trap 132 is configured as from flow region 106/ channel 122 of domain receives or captures multiple speck bodies.In some cases, trap 132 includes and single target speck body The roughly equal volume of volume.

Trap 132 may further include opening, and opening is configured to assist in target speck body and flows into trap 132. In some cases, trap 132 includes with the height and width being sized according to the size of single target speck body Thus opening prevents other speck bodies (or larger-sized speck body) from entering speck body trap.Trap 132 can be into one Step includes other features for being configured as helping to be maintained at target speck body in trap 132.In some cases, trap 132 completely cut off the aligned on opposite sides in opening and channel 122 of fence and disposed thereon relative to microfluid so that around being parallel to When the axis in channel 122 tilts microfluidic device 100, the speck body that is captured is according to making speck body fall into opening for isolation rail Leave trap 132 in track in mouthful.In some cases, trap 132 includes the wing passage 134 less than target speck body, In order to by the flowing of trap 132, to increase the possibility for capturing speck body in trap 132.

As discussed in more detail below, in some embodiments, electrowetting (EW) power via one or more electrodes (not Show) at one or more positions on the surface of the support construction 104 (and/or lid 110) that is applied to microfluidic device 100 (for example, position in flow region and/or isolation fence), is located at microfluidic circuit 120 with manipulation, transmission, separation and sorting In drop.For example, in some embodiments, EW power is applied in one on the surface of support construction 104 (and/or lid 110) At a or more position, drop is transferred to from flow region 106 in desired microfluid isolation fence.In some implementations In example, EW power be used to prevent the drop completely cut off in fence (such as isolation fence 124,126,128 or 130) from wherein shifting. In addition, in some embodiments, EW power, which is selectively removed from isolation fence for introduction according to the present invention, had previously collected Drop.In some embodiments, EW power includes photoelectricity wetting (OEW) power,

In some embodiments, dielectrophoresis (DEP) power is applied to fluid by one or more electrode (not shown) On medium 180 (for example, in flow region and/or isolation fence), it is located therein with manipulation, transmission, separation and sorting micro- Object.For example, in some embodiments, DEP power is applied in one or more parts of microfluidic circuit 120, will be single A speck body is transferred to from flow region 106 in desired microfluid isolation fence.In some embodiments, come using DEP power Prevent speck body in isolation fence (such as isolation fence 124,126,128 or 130) from wherein shifting.In addition, in some realities It applies in example, the speck body previously collected is removed from isolation fence come selectively introduction according to the present invention using DEP power.One In a little embodiments, DEP power includes photoelectron tweezers (OET) power.

In some embodiments, DEP and/or EW power is combined with other power of such as flowing and/or gravity etc, with behaviour Speck body and/or drop in vertical, transmission, separation and sorting microfluidic circuit 120.For example, capsule 102 can tilt (such as Pass through reclining device 190), 106/ channel 122 of flow region and the speck body being located therein are located in microfluid isolation and enclosed Above column, and gravity can will be in speck body and/or droplet transport to fence.It in some embodiments, can be in other power Apply DEP and/or EW power before.It in other embodiments, can be in the after-applied DEP and/or EW power of other power.In other feelings Under condition, DEP and/or EW power can replace application simultaneously or with other power with other power.

Microfluidic device power configuration.As described above, the control of system and monitoring device may include in microfluid The power plant module of selection and the mobile such as object of speck body or drop in the microfluidic circuit of equipment.Depending on by mobile object The type of body and other Considerations, microfluidic device of the invention can have a variety of power configurations.Particularly, microfluidic device 100 support construction 104 and/or lid 110 may include electrowetting (EW) configuration, for selectively inducing microfluidic circuit The EW power on the drop in fluid media (medium) 180 in 120, to select, capture and/or move individual drop or drop group. In certain embodiments, microfluidic device of the invention can include with the EW first parts configured and with dielectrophoresis (DEP) second part configured.Therefore, the support construction 104 of microfluidic device 100 and/or at least part of lid 110 can be with It is configured including DEP, DEP power is induced on the speck body in the selectively fluid media (medium) 180 in microfluidic circuit 120, To select, capture and/or move individual speck body or micro- group of objects.

Electrowetting configures.In certain embodiments, microfluidic device of the invention may include electrowetting configuration comprising Substrate with dielectric layer and drop actuation surfaces, drop actuation surfaces include being covalently bound to the hydrophobic layer of dielectric layer.Dielectric Layer can be located at below hydrophobic layer so that the drop on substrate is in direct contact hydrophobic layer.Fig. 2A shows that this microfluid is set The example of a standby part.

As shown, device 400 may include base portion 104, base portion 104 includes substrate and at least one electrode (for example, the One electrode) 418.Substrate may include various layers, including outer hydrophobic layer 412, inner-dielectric-ayer 414, conductive layer 416,418 and of electrode Optional support element 420.Hydrophobic layer 412 and inner-dielectric-ayer 414 can provide the inside table for the substrate 102 for partly limiting capsule Face.

Device 400 further includes lid 110, and lid 110 includes outer hydrophobic layer 422, may include the internal layer 428 of at least one electrode And optional support element 430.Lid 110 and base portion 104 are substantially parallel to each other and by spacer element 108 (for example, miniflow Body return path materials) it is bonded together, to limit the capsule 435 for being configured to accommodate liquid medium.Liquid medium can be for example Hydrophobic liquid, such as oil.In addition, capsule 435 can accommodate the drop of liquid 440, such as aqueous medium.Typically, liquid The liquid of medium and drop is selected as not miscible liquid.

Spacer element 108 may include polymer.The polymer can be such as organic polymer based on silicon, such as poly- Dimethyl siloxane (PDMS) or photo-patterned polysiloxanes (PPS), can both obtain from DOW CORNING.Optionally, Every element 108 may include epoxy adhesive.Epoxy adhesive can be the material of such as SU-8 or equivalent type.Interval Element 108 can at least 30,40,50,60,70,80,90, the thickness of 100 or more microns is (that is, the interior table of substrate 104 Gap between face and lid 110).Thus, for example, the thickness of spacer element 108 can be 30-60 microns, 40-80 microns, 50- 100 microns, 60-120 microns, 70-140 microns, 75-150 microns, 80-160 microns, 90-180 microns or 100-200 microns.

Spacer element 108 can limit one or more microfluidic channels in capsule.In addition, spacer element 108 can Further to limit multiple chambers (or isolation fence) in capsule, wherein each chamber fluid be connected at least one miniflow Body channel and from wherein opening.Thus, for example, spacer element 108 can limit single microfluidic channel and connect with its fluid The multiple chambers connect or multiple microfluidic channels, each channel are fluidly connected to multiple chambers.In addition, each chamber can be with It is fluidly coupled to more than one microfluidic channel, as shown in Figures 6 and 7.

(do not show when at least one electrode 418 of substrate 104 and at least one electrode 428 of lid 110 are connected to AC voltage sources Go out) opposite terminal when, electrowetting power can be applied to the outer hydrophobic surface 412 with substrate 104 (that is, drop by substrate 104 Actuation surfaces) contact aqueous drop.In certain embodiments, be used in microfluidic device realize drop based on electrowetting Movement AC voltages be at least 20 volts of peak-to-peak voltages (ppV) (for example, about 20 to 80ppV, about 20 to 60ppV, about 25 to 50ppV, about 25 to 40ppV or about 25 to 35ppV).In certain embodiments, it is used to realize the base of drop in microfluidic device In the frequency of the AC voltages of the movement of electrowetting be about 1 to 100kHz (for example, about 5 to 90kHz, about 10 to 80kHz, about 15 to 70kHz, about 20 to 60kHz, about 25 to 50kHz or about 30 to 40kHz).

The outer hydrophobic layer 412 of substrate 104 and the outer hydrophobic layer 422 of lid 110 can include respectively being covalently bound to substrate respectively The closelypacked self-association molecular monolayer of 104 inner-dielectric-ayer 414 or the internal layer 428 of lid 110.The self-association molecule of single layer Including enough two-dimentional bulk densities, to generate hydrophobic barrier between the surface that single layer is combined and hydrophilic liquid (that is, prevent polar molecule or other chemical substances be embedded in and/or penetrate into single layer).The accumulation of the single layer of dense accumulation is close Degree will depend on used self-association molecule.Dense accumulation single layer including alkyl-blocked siloxanes is typically included at least 1x10¹⁴Molecule/cm²(for example, at least 1.5x10¹⁴、2.0x10¹⁴、2.5x10¹⁴Or more molecule/cm²)。

As described in greater detail below, self-association molecule can include respectively the link group, such as siloxane group or Phosphonyl group.Siloxane group can be covalently bound to the molecule of inner-dielectric-ayer 414 or internal layer 428.Similarly, phosphonyl group can It is covalently bound to the molecule of inner-dielectric-ayer 414 or internal layer 428.Self-association molecule can include long chain hydrocarbons, can be unbranched 's.Therefore, self-association molecule may include alkyl-blocked siloxanes or alkyl-blocked phosphonic acids.Long chain hydrocarbons can include at least The chain of 10 carbon (for example, at least 16,18,20,22 or more carbon).Self-association molecule can include fluorocarbons Chain.Thus, for example, self-association molecule can include the siloxanes of fluoro-alkyl sealing end or the phosphonic acids of fluoro-alkyl sealing end.Fluorination Carbochain can have chemical formula CF₃-(CF₂)m-(CH₂) n-, wherein m is at least 2, n 0,1, two or more, and m+n is at least 9。

Single layer self-association molecule, which can have, to be less than about 5 nanometers (for example, about 1.0 to about 4.0 nanometers, about 1.5 to about 3.0 receives Rice or about 2.0 to about 2.5 nanometers) thickness.

The outer hydrophobic layer 412 of substrate 104 can be patterned such that compared with the rest part of outer hydrophobic layer, select area Domain relative hydrophobic.For example, this can by by the voltage drop on following inner-dielectric-ayer 122 increase to 50ppV or higher (such as 60,65,70,75,80 or more ppV) a period of time is reached to realize.It is without being bound by theory, it is believed that relative hydropathic region includes embedding Enter the hydrone in single layer.

In some embodiments, the inner-dielectric-ayer of substrate may include one or more oxide skin(coating)s.For example, interior dielectric Layer may include single oxide skin(coating) (such as metal oxide layer) or be made from it.Alternatively, inner-dielectric-ayer may include two layers Or it is formed by two layers.In some embodiments, one layer can be silica or silicon nitride, and another layer can be metal oxidation Object, such as aluminium oxide.In certain embodiments, the thickness of metal oxide layer can be in about 15nm to about 45nm or about 30nm To about 40nm or about 33nm to about 36nm.Atomic layer deposition (ALD) technology depositing metal oxide can be passed through Layer, and can be deposited comprising silica or silicon nitride by plasma enhanced chemical vapor deposition (PECVD) technology Layer.

In yet another embodiment, inner-dielectric-ayer may include three layers of dielectric material.In some embodiments, first layer can With including the metal oxide that can be clipped between silicon dioxide layer and silicon nitride layer, such as aluminium oxide, hafnium oxide etc..Certain In embodiment, the thickness of metal oxide layer can be in the range of about 5nm to about 20nm, and the layer can pass through atomic layer (ALD) technology of deposition deposition.Silicon oxide layer can also be by ALD deposition, and can have the thickness of about 2nm to about 10nm. Silicon nitride layer can be deposited by plasma enhanced chemical vapor deposition (PECVD) technology and can have about 80nm to about The thickness of 100nm or about 90nm thickness.

Regardless of constituting the number of plies of inner-dielectric-ayer, inner-dielectric-ayer can be with about 50 to 105 nanometers of thickness and/or about 50 To 150 kilohms of impedance, preferred embodiment is about 100 kilohms.

Substrate 104 may include the photoresponsive layer 146 of the first side with contact inner-dielectric-ayer 414.Photoresponsive layer 416 The second side can contact at least one electrode 418.Photoresponsive layer 416 can include amorphous silicon hydride (a-Si：H).For example, a- Si：H can include about 8% to 40% hydrogen (that is, being calculated as quantity/hydrogen atom of 100* hydrogen atoms and the sum of silicon atom). a-Si：H layers can have at least about 500 nanometers (for example, at least about 600 to 1400, about 700 to 1300, about 800 to 1200, about 900 to 1100 or about 1000 nanometers) thickness.However, a-Si：H layers of thickness can become according to the thickness of inner-dielectric-ayer 414 Change, so as to the reality when substrate 104 be in open state (that is, illuminated and conducting) and closed state (that is, dark be not turned on) The impedance of existing inner-dielectric-ayer 414 and a-Si：Appropriate difference between H layers of impedance.For example, the impedance of inner-dielectric-ayer 414 can be with It is tuned to about 50kOhm to about 150kOhm, and a-Si：H layers of impedance can in the off state be adjusted at least about 0.5MOhm, and it is less than or equal to about 1kOhm in the on state.These are only example, but they show impedance How to be tuned to realize photoresponse (in this case, the photoconductive) layer 416 for the ON/OFF performance for showing robust.In substrate 104 have by a-Si：In the embodiment of the photoresponsive layer 416 of H layers of formation, substrate 104 can be optionally included positioned at photoresponse Floating electrode pad between layer 416 and inner-dielectric-ayer 414.Such as this has been described in No. 6,958,132 United States Patent (USP) Kind floating electrode pad, content are incorporated herein by reference.

Alternatively, photoresponsive layer 416 may include multiple conductors, each conductor controllably connects via phototransistor switch It is connected at least one electrode of substrate 102.The conductor controlled by phototransistor switch is well known in the present art, and Through being described in such as No. 2014/0124370 U.S. Patent application, content is incorporated herein by reference.

Substrate 104 may include the single electrode 418 for being configured to connect to AC voltage sources.Single electrode 418 may include oxygen Change indium tin (ITO) layer, such as can be formed in glass supports 420.Alternatively, single electrode 418 may include conductive silicon layer. In other embodiments, substrate 104 may include separately addressable multiple electrodes, EWOD devices as known in the art Mode.Separately addressable electrode can be connected to one or more AC voltage sources by corresponding transistor switch.

Lid 110 can be further comprised and 422 juxtaposed dielectric layer (not shown) of hydrophobic layer and juxtaposition in a manner of substrate Conductive layer (not shown) between dielectric layer and electrode 428.Therefore, microfluidic device 400 can make substrate 104 and lid 110 The two is configured to provide electrowetting power to the aqueous drop 440 in capsule 435.In such embodiments, lid 110 Dielectric layer can by the inner-dielectric-ayer 414 disclosed herein for substrate 104 it is any in a manner of configure, and lid 104 Conductive layer can by the conductive layer 126 disclosed herein for substrate 102 it is any in a manner of configure.

Dielectrophoresis (DEP) configures.As discussed herein, microfluidic device of the invention may include with DEP configurations Part.Another example is microfluidic devices 200 shown in Fig. 1 C and 1D for the part.Although for simplicity, Fig. 1 C and The vertical cross-section that 1D respectively illustrates a part for the capsule 102 of the microfluidic device 200 with open area/chamber 202 regards Figure and horizontal sectional view, it should be appreciated that region/chamber 202 can be the fluid circuit element for having more detailed structure A part, such as growth chamber, isolation fence, flow region or flow channel.In addition, microfluidic device 200 may include Other fluid circuit elements.For example, microfluidic device 200 may include multiple growth chambers or isolation fence and/or one or More flow regions or flow channel, for example, herein in regard to microfluidic device 100 it is described those.DEP configurations can tie In any this fluid circuit element or its selected section for closing microfluidic device 200.It is to be further understood that any Microfluidic device component and system unit above-mentioned or described below can be incorporated to microfluidic device 200 and/or be set with microfluid Standby 200 are used in combination.E.g., including the system 150 of above-mentioned control and monitoring device 152 can with microfluidic device 200 (including It is one or more in medium module 160, power plant module 162, image-forming module 164, tilt module 166 and other modules 168) It is used together.

As shown in Figure 1 C, microfluidic device 200 includes the electrode active for having bottom electrode 204 and covering bottom electrode 204 Change the support construction 104 of substrate 206 and the lid 110 with top electrodes 210, wherein top electrodes 210 and bottom electrode 204 are spaced apart.The apparent surface of top electrodes 210 and electrode activation substrate 206 limited areas/chamber 202.Included in region/ Therefore medium 180 in chamber 202 provides resistance connection between top electrodes 210 and electrode activation substrate 206.It also shows It is configured to connect to bottom electrode 204 and top electrodes 210 and generates the power supply 212 of bias voltage between the electrodes, such as exist It is generated as needed for DEP power in region/chamber 202.Power supply 212 can be such as exchange (AC) power supply.

In certain embodiments, the DEP configurations that microfluidic device 200 shown in Fig. 1 C and 1D can be with optical actuation. Therefore, the change pattern for the light 218 from light source 216 that can be controlled by power plant module 162 can selectively activate and stop With the change pattern of the DEP electrodes at the region 214 of the inner surface 208 of electrode activation substrate 206.(hereinafter, there is DEP The region 214 of the microfluidic device of configuration is referred to as " DEP electrode zones ".) as shown in figure iD, it is directed to electrode activation substrate Light pattern 218 on 206 inner surface 208 can be irradiated with such as square pattern selection DEP electrode zones 214a (with White displays).Not illuminated DEP electrode zones 214 (intersecting hachure) are known as " dark " DEP electrode zones 214 below.Pass through DEP electrode activations substrate 206 is (that is, from bottom electrode 204 until electrode activation substrate 206 and the medium in flow region 106 180 have a common boundary inner surfaces 208) relative impedances be more than at each dark DEP electrode zones 214 by region/chamber 202 The relative impedances of medium 180 (that is, from inner surface 208 of electrode activation substrate 206 to the top electrodes 210 of lid 110).However, Illuminated DEP electrode zones 214a shows the relative impedances of the reduction by electrode activation substrate 206, and the relative impedances are small In the relative impedances for passing through the medium 180 in region/chamber 202 at each illuminated DEP electrode zones 214a.

In the case where activating power supply 212, above-mentioned DEP configuration is in illuminated DEP electrode zones 214a and adjacent dark Electric-force gradient is generated in fluid media (medium) 180 between DEP electrode zones 214, this is generated again in attraction or repelling fluid medium 180 Near speck body (not shown) local DEP power.Therefore, can the inner surface 208 of region/chamber 202 it is many not It is selected by changing light pattern 218 projected from light source 216 in microfluidic device 200 at same this DEP electrode zones 214 Activate and deactivate to selecting property the DEP electrodes of the speck body attracted or in repelling fluid medium 180.DEP power is to attract or repel Neighbouring speck body may depend on the dielectric property etc. of the frequency and medium 180 and/or speck body (not shown) of power supply 212 Parameter.

The square pattern 220 of illuminated DEP electrode zones 214a is only example shown in Fig. 1 C.Throwing can be passed through The light pattern 218 being mapped in equipment 200 irradiates any pattern of (and thus activate) DEP electrode zones 214, and irradiate/ The pattern of the DEP electrode zones 214 of activation can repeat to change by changing or moving light pattern 218.

In some embodiments, electrode activation substrate 206 may include photoconductive material or is made of photoconductive material. In such embodiment, the inner surface 208 of electrode activation substrate 206 can be undistinguishable.For example, electrode activation substrate 206 May include amorphous silicon hydride (a-Si：H it) layer or is made from it.a-Si：H (can be pressed comprising for example, about 8% to 40% hydrogen The sum of 100* numbers of hydrogen atoms/hydrogen atom and silicon atom calculates).a-Si：H layers can be with about 500nm to about 2.0 μm of thickness. In such embodiments, according to light pattern 218, DEP electrode zones 214 can be in the inner surface 208 of electrode activation substrate 206 It is formed with any pattern from anywhere in upper.Therefore, the quantity of DEP electrode zones 214 and pattern need not be fixed, but It can correspond to light pattern 218.Such as in No. 44,711 United States Patent (USP)s (Wu et al.) of RE (initially as No. 7,612,355 U.S. Patent Publication) in have been described with including that the microfluidic devices that configure of DEP of photoconductive layer as described above show Example, entire contents are incorporated herein by reference.

In other embodiments, electrode activation substrate 206 may include substrate, the substrate include multiple doped layers, electricity absolutely Edge layer (or region) and the conductive layer for forming semiconductor integrated circuit, it is known such as in semiconductor applications.For example, electrode active It may include multiple phototransistors to change substrate 206, such as including lateral bipolar phototransistor, and each phototransistor corresponds to In DEP electrode zones 214.Alternatively, electrode activation substrate 206 may include controlled by phototransistor switch electrode (for example, Conductive metal electrode), each such electrode corresponds to DEP electrode zones 214.Electrode activation substrate 206 may include this The pattern of phototransistor or phototransistor coordination electrode.For example, the pattern can be arranged in rows and columns substantially just The array of rectangular phototransistor or phototransistor coordination electrode, as shown in Figure 2 B.Alternatively, the pattern can form six The phototransistor of the substantially hexagon of square lattice or the array of phototransistor coordination electrode.Regardless of pattern, circuit Element can be in the DEP electrode zones 214 at the inner surface 208 of electrode activation substrate 206 and formation electricity between bottom electrode 210 Connection, and those electrical connections (that is, phototransistor or electrode) can be selectively activated and deactivated by light pattern 218.When When not being activated, each electrical connection can have high impedance so that by electrode activation substrate 206 (that is, from bottom electrode 204 To the inner surface 208 of the electrode activation substrate 206 with 180 interfaces of medium in region/chamber 202) relative impedances be more than By medium 180 (that is, from the inner surface 208 of electrode activation substrate 206 to the top of lid 110 at corresponding DEP electrode zones 214 Portion's electrode 210) relative impedances.However, when by photoactivation in light pattern 218, pass through the opposite of electrode activation substrate 206 Impedance is less than at each illuminated DEP electrode zones 214 through the relative impedances of medium 180, to activate corresponding DEP electricity DEP electrodes at polar region domain, as described above.It therefore, can be in region/chamber 202 in a manner of being determined by light pattern 218 Electrode activation substrate 206 inner surface 208 at many different DEP electrode zones 214 at selectively activate and deactivate Attract or repel the DEP electrodes of the speck body (not shown) in medium 180.

It is had been described in such as No. 7,956,339 United States Patent (USP) (Ohta et al.) with including phototransistor The example (see, for example, equipment 300 shown in Figure 21 and Figure 22 and its description) of the microfluidic device of electrode activation substrate, Full content is incorporated herein by reference.With the micro- of the electrode activation substrate for including the electrode controlled by phototransistor switch The example of fluid device described in the United States Patent (USP) (Short et al.) of such as Publication No. 2014/0124370 (referring to Such as equipment 200,400,500,600 and 900 shown in entire attached drawing and its description), entire contents are incorporated by reference into this Text.

DEP configuration microfluidic device some embodiments in, top electrodes 210 be capsule 102 the first wall (or lid 110) a part, and electrode activation substrate 206 and bottom electrode 204 are the parts of the second wall (or support construction 104). Region/chamber 202 can be between the first wall and the second wall.In other embodiments, electrode 210 be the second wall (or support knot Structure 104) a part, and one or both of electrode activation substrate 206 and/or electrode 210 are the first wall (or lids 110) A part.In addition, light source 216 can be alternatively for irradiation capsule 102 from below.

Using microfluidic devices 200 of Fig. 1 C to Fig. 1 D with DEP configurations, power plant module 162 can be by by light figure Case 218 projects the speck body (not shown) come in equipment 200 in the medium in selection region/chamber 202 180, with around And capture the DEP electricity of the inner surface 208 of active electrode activation substrate 206 in the pattern (for example, square pattern 220) of speck body First group of one or more DEP electrode at the 214a of polar region domain.Then, power plant module 162 can be by relative to equipment 200 Mobile light pattern 218 come move the speck body of capture with activate second group at DEP electrode zones 214 it is one or more DEP electrodes.Alternatively, equipment 200 can be moved relative to light pattern 218.

In other embodiments, microfluidic device 200 can have independent of the inner surface in electrode activation substrate 206 The DEP of the photoactivation of DEP electrodes at 208 is configured.For example, electrode activation substrate 206 may include and include at least one electricity The selective addressable of surface (for example, lid 110) relative positioning of pole and electrode can be encouraged.Switch is (for example, in semiconductor substrate Transistor switch) can selectively open and close, to activate or be passivated the DEP electrodes at DEP electrode zones 214, from And generate net DEP power on the speck body (not shown) in region/chamber 202 near the DEP electrodes of activation.Depending on such as Such characteristic of the dielectric property of speck body in the frequency and medium (not shown) and/or region/chamber 202 of power supply 212, DEP power can attract or repel neighbouring speck body.By selectively activating and deactivating groups of DEP electrodes (for example, in shape At the groups of DEP electrode zones 214 of square pattern 220), one or more speck bodies in region/chamber 202 can It is captured and is moved in region/chamber 202.Power plant module 162 in Figure 1A can control such switch and therefore activate With the individual DEP electrodes in deactivated DEP electrodes to select, trap and move the specific speck body of encircled area/chamber 202 (not shown).With being in the art including selective addressable and microfluidic device that the DEP of electrode can be encouraged to configure Know, and has been retouched in the 6th, 294, No. 063 (Becker etc.) and the 6th, 942, No. 776 (Medoro) United States Patent (USP) It states, entire contents are incorporated herein by reference.

The microfluidic device configured with electrowetting and dielectrophoresis (DEP).Fig. 4 is according to various embodiments integrated multiple The microfluidic device of microfluidic applications or the vertical cross-section diagram of device 450.Equipment 450 includes two different parts (although can It is more to have), each part has the configuration of single microfluid.Part 460 is configured including electrowetting comprising includes the base of substrate Portion 104.Substrate includes various layers, including outer hydrophobic layer 412, inner-dielectric-ayer 414, conductive layer 416 and electrode 418.Hydrophobic layer 412 With inner-dielectric-ayer 414 can provide partly limit capsule 435 substrate towards inner surface.Part 460 further includes comprising electricity The lid 110 of pole 428 and outer hydrophobic layer 422 and the microfluidic circuit material 108 for connecting base portion 104 with lid 110, microfluid Return path materials 108 additionally aid the microfluidic circuit for limiting electrowetting part, including are configured as accommodating not miscible fluid Capsule 435.

The part 470 of microfluidic device 450 includes dielectrophoresis DEP configurations comprising base portion 104, first electrode 479, electricity Pole activates substrate 474 and partly limits the inward-facing surface of capsule 475.Part 470 further includes the lid for including electrode 468 110 and base portion 104 is connect to lid 110 and is additionally aided the microfluidic circuit material of the microfluidic circuit for limiting DEP parts Material 108.

As shown in figure 4, electrowetting part 460 and the parts DEP 470 can share identical base portion 104 and lid 110, and it Substrate and electrode do not share.The electrowetting part 460 and the parts DEP 470 of equipment 450 can be engaged by bridge 465, bridge 465 can be that pipeline, jointing material wait or any combination thereof.

Fig. 5 is vertical section of the microfluidic device for integrating multiple microfluidic applications or device 500 according to various embodiments Face figure.As equipment 400, equipment 500 includes two different parts (although can have more), and each part has single Microfluid configures.Specifically, part 460 is configured including electrowetting, part 470 is configured including DEP.Such as corresponding reference numeral Shown, the various parts of equipment 500 have part corresponding with the component in equipment 400.However, equipment 500 has monolithic Substrate, which has conductive layer 416, first electrode 418 and second electrode 428, all these all by 460 He of part 470 share.

Figure 19 A and 19B are provided according to the embodiment for combining Fig. 5 descriptions for the electrical addressing operation in terms of function The view of expression.Described in conjunction with Figure 5 as before, two microfluidic procedures of the system integration such as share one-piece substrate 416 DEP and EW modules it is discribed.In this embodiment, DEP (it can be OET) modules have lower relative to EW modules Impedance.During operation, the impedance of EW modules overcomes the impedance of DEP modules, and substantially makes DEP module short circuits.

In one embodiment as shown in Figure 19 A, OEP modules with the frequency within the scope of 100kHz to 10mHz by being applied The voltage in 1-10 volt ranges is added to operate.In the same embodiment, as shown in Figure 19 B, OEW modules by with 1kHz extremely Frequency within the scope of 300kHz applies the voltage in 10-100 volt ranges to operate.In a preferred embodiment, OEP modules are logical It crosses and 5 volts of voltage is applied to operate with the frequency of 1MHz, and OEW modules pass through the voltage of 30 volts of the frequency application with 30kHz To operate.

Completely cut off fence.General isolation fence 224,226 and 228 is shown in the microfluidic device 230 shown in Fig. 2A -2C Non-limiting example.Each isolation fence 224,226 and 228 may include limiting the isolation structure 232 of isolated area 240 and inciting somebody to action Isolated area 240 is fluidly connected to the bonding pad 236 in channel 122.Bonding pad 236 may include the proximal open for leading to channel 122 234 and lead to the distal openings 238 of isolated area 240.Bonding pad 236, which can be configured so that from channel 122, flows into isolation fence 224, the maximum penetration of the flowing of 226,228 fluid media (medium) (not shown) does not extend in isolated area 240.Therefore, by In bonding pad 236, speck body (not shown) or other materials in the isolated area 240 of isolation fence 224,226,228 are set (not shown) therefore can be with the flow separation of the medium 180 in channel 122 and substantially unaffected.

The isolation fence 224,226 and 228 of Fig. 2A -2C respectively has the single opening for being directly communicated to channel 122.Isolation is enclosed The opening on column is laterally-opening from channel 122.Electrode activation substrate 206 is located at channel 122 and 224,226 and 228 liang of fence of isolation Below person.The upper surface setting of electrode activation substrate 206 in the capsule for the isolation fence for forming the bottom surface of isolation fence exists Formed microfluidic device flow channel (or flow region) bottom surface channel 122 (or if channel be not present if for stream Dynamic region) in electrode activation substrate 206 upper surface identical highly or substantially upper identical height at.Electrode activation substrate 206 can be undistinguishable or can have an irregular or patterned surface, and the surface is from its maximum height to minimum Back variation be less than about 3 microns, 2.5 microns, 2 microns, 1.5 microns, 1 micron, 0.9 micron, 0.5 micron, 0.4 micron, it is 0.2 micro- Rice, 0.1 micron or smaller.Pass through the change of the height in the upper surface of channel 122 (or flow region) and the substrate for completely cutting off fence Change be smaller than isolation fence wall or microfluidic device wall height about 3%, 2%, 1%, 0.9%, 0.8%, 0.5%, 0.3% or 0.1%.Although microfluidic device 200 is described in detail, this is also applied for any microfluidic device as described herein 100、230、250、280、290、600、700。

Therefore channel 122 can be the example in the region being swept, and completely cut off the isolated area of fence 224,226,228 240 can be the example in the region not being swept.As described above, channel 122 and isolation fence 224,226,228 can be configured To include one or more fluid media (medium)s 180.In the example shown in Fig. 2A -2B, port 222 be connected to channel 122 and Fluid media (medium) 180 is allowed to be introduced into microfluidic device 230 or removed from microfluidic device 230.Introduce fluid media (medium) 180 it Before, microfluidic device can use the gas of such as carbon dioxide gas to load.Once microfluidic device 230 includes fluid media (medium) 180, the flowing 242 of the fluid media (medium) 180 in channel 122 selectively can be generated and be stopped.For example, as shown, port 222 can be arranged at the different location (for example, opposite end) in channel 122, and can be held from one as entrance Mouth 222 creates the flowing 242 of medium to another port 222 used for export.

Fig. 2 C show the exemplary detailed view of isolation fence 224 according to the present invention.Also show speck body 246 Example.

As it is known, fluid media (medium) 180 passes through the stream of the proximal open 234 of isolation fence 224 in microfluidic channel 122 Dynamic 242 can cause medium 180 to enter and/or leave the secondary flow 244 of isolation fence 224.In order to will completely cut off fence 224 every It is isolated with secondary flow 244 from the speck body 246 in area 240, completely cuts off the length L of the bonding pad 236 of fence 224_con(that is, from close Side opening 234 is to distal openings 238) penetration depth D that secondary flow 244 enters bonding pad 236 should be greater than_p.Secondary Flow Dynamic 244 penetration depth D_pConfiguration depending on the speed of fluid media (medium) 180 flowed in channel 122 and with channel 122 The 234 related various parameters of proximal open in channel 122 are arrived with bonding pad 236.For given microfluidic device, channel 122 Configuration with opening 234 will be fixed, and the rate of the flowing 242 of the fluid media (medium) 180 in channel 122 will be variable. Therefore, for each isolation fence 224, the maximum speed of the flowing 242 of the fluid media (medium) 180 in channel 122 can be identified V_max, ensure the penetration depth D of secondary flow 244_pNo more than the length L of bonding pad 236_con.As long as the fluid in channel 122 The rate of the flowing 242 of medium 180 is no more than maximum speed V_max, then caused by secondary flow 244 can be limited to channel It 122 and bonding pad 236 and is maintained at except isolated area 240.Therefore, flowing 242 of the medium 180 in channel 122 will not incite somebody to action Speck body 246 pulls out isolated area 240.On the contrary, the speck body 246 in isolated area 240 will stay in isolated area 240, and It is unrelated with the flowing 242 of fluid media (medium) 180 in channel 122.

Moreover, as long as the rate of the flowing 242 of medium 180 in channel 122 is no more than V_max, the fluid in channel 122 is situated between The flowing 242 of matter 180 will not be such that miscellaneous granules (for example, micron particles and/or nano particle) are moved into from channel 122 Completely cut off the isolated area 240 of fence 224.Therefore, the length L of bonding pad 236_conMore than the maximum penetration D of secondary flow 244_p It can prevent the miscellaneous granules pair one from channel 122 or another isolation fence (such as isolation fence 226,228 in Fig. 2 D) The pollution of a isolation fence 224.

Because channel 122 and the bonding pad 236 of isolation fence 224,226,228 may in by channel 122 medium 180 Flowing 242 influences, so channel 122 and bonding pad 236 are considered being swept of microfluidic device 230 (or flowing) area Domain.On the other hand, the isolated area 240 of isolation fence 224,226,228, which is considered, is not swept (or non-current) region. For example, the component (not shown) in channel 122 in first fluid medium 180 can be substantially only through the component of first medium 180 Diffusion from channel 122 by bonding pad 236 and enter isolated area 240 in second fluid medium 248 by in isolated area 240 Second fluid medium 248 mix.Similarly, the component (not shown) of the second medium 248 in isolated area 240 can be substantially First in bonding pad 236 and entrance channel 122 is only passed through from isolated area 240 by the diffusion of the component of second medium 248 Medium 180 and mixed with the first medium 180 in channel 122.In some embodiments, completely cut off the isolated area and flow region of fence Between by spread carry out fluid media (medium) exchange degree be more than fluid communication about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or greater than about 99%.First medium 180 can be medium identical with second medium 248 or not Same medium.In addition, first medium 180 and second medium 248 can be initially identical, then become difference (for example, passing through Adjusting of one or more cells to second medium 248 in isolated area 240, or flow through Jie in channel 122 by changing Matter 180).

As described above, the maximum penetration of secondary flow 244 caused by flowing 242 by the fluid media (medium) 180 in channel 122 Depth D_pIt can depend on multiple parameters.The example of such parameter includes：The shape in channel 122 is (for example, channel can will be situated between Matter is directed in bonding pad 236, so that medium is transferred out of bonding pad 236, or in the bonding pad substantially perpendicular to channel 122 Medium is directed in channel 122 on the direction of 236 proximal open 234)；Width W of the channel 122 at proximal open 234_ch (or sectional area)；With the width W of the bonding pad 236 at proximal open 234_con(or sectional area)；Fluid media (medium) 180 is in channel The speed V of flowing 242 in 122；The viscosity etc. of first medium 180 and/or second medium 248.

In some embodiments, the size of channel 122 and isolation fence 224,226,228 can be relative in channel 122 The vector of the flowing 242 of fluid media (medium) 180 orients as follows：Channel width W_ch(or the sectional area in channel 122) can substantially hang down Directly in the flowing 242 of medium 180；Width W of the bonding pad 236 at opening 234_con(or sectional area) may be substantially parallel to lead to The flowing 242 of medium 180 in road 122；And/or the length L of bonding pad_conIt may be substantially perpendicular to the medium in channel 122 180 flowing 242.It the above is only example, and the relative position of channel 122 and isolation fence 224,226,228 can be opposite In each other in other orientation.

As shown in Figure 2 C, the width W of bonding pad 236_conCan proximally be open 234 to distal openings 238 be uniform. Therefore, the width W of the bonding pad 236 at distal openings 238_conCan be herein be bonding pad 236 at proximal open 234 Width W_conIn any range identified.Alternatively, the width W of the bonding pad 236 at distal openings 238_conIt can be more than close The width W of bonding pad 236 at side opening 234_con。

As shown in Figure 2 C, the width of the isolated area 240 at distal openings 238 can be with the bonding pad at proximal open 234 236 width W_conIt is essentially identical.Therefore, the width of the isolated area 240 at distal openings 238 can be herein for bonding pad The 236 width W at proximal open 234_conIn any range identified.Alternatively, isolated area 240 at distal openings 238 Width can be more than or less than the width W of the bonding pad 236 at proximal open 234_con.Moreover, distal openings 238 can be less than Proximal open 234, and the width W of bonding pad 236_conIt can narrow between proximal open 234 and distal openings 238.Example Such as, bonding pad 236 can be using a variety of different geometries (such as chamfering bonding pad, chamfer bonding pad) and in proximal open Narrow between distal openings.In addition, any part or subdivision of bonding pad 236 are (for example, bonding pad and proximal open 234 Adjacent portion) it can narrow.

Fig. 2 D-2F depict the microfluidic device 250 comprising microfluidic circuit 262 and flow channel 264 another show Example property embodiment, is the modification of the corresponding microfluidic device 100, circuit 132 and channel 134 of Fig. 1.Microfluidic device 250 is also With multiple isolation fences 266, the isolation fence 266 is above-mentioned isolation fence 124,126,128,130,224,226 or 228 Additional variations.Particularly, it should be understood that the isolation fence 266 of equipment 250 shown in Fig. 2 D-2F can replace setting Any of above isolation fence 124,126,128,130,224,226 or 228 in standby 100,200,230,280,290 or 320.Class As, microfluidic device 250 is another modification of microfluidic device 100, and can also be had and above-mentioned microfluidic device 100,200,230,280,290,320 identical or different DEP configurations and any other microfluidic system portion described herein Part.

The microfluidic device 250 of Fig. 2 D-2F includes that support construction is (invisible in Fig. 2 D-2F, but can be retouched with Figure 1A The support construction 104 for the equipment 100 painted is identical or substantially similar), microfluidic circuit structure 256 and lid (can not in Fig. 2 D-2F See, but can be identical or substantially similar as the lid 122 of equipment 100 described in Figure 1A).Microfluidic circuit structure 256 includes frame Frame 252 and microfluidic circuit material 260, can be with the frame 114 and microfluidic circuit material of equipment shown in Figure 1A 100 116 is identical or substantially similar.As shown in Figure 2 D, the microfluidic circuit 262 limited by microfluidic circuit material 260 may include multiple Multiple channels 264 (show two, but can have more) that isolation fence 266 is fluidly connected to.

Each isolation fence 266 may include isolation structure 272, the isolated area 270 in isolation structure 272 and bonding pad 268.From distal openings 276 of the proximal open 274 from channel 264 at isolation structure 272, channel 264 is flowed in bonding pad 268 Body is connected to isolated area 270.In general, discussed above, the stream of the first fluid medium 254 in channel 264 of B and 2C according to fig. 2 Dynamic 278 can generate two that first medium 254 enters from channel 264 and/or leave the corresponding bonding pad 268 of isolation fence 266 Secondary flowing 282.

As shown in Figure 2 E, each the bonding pad 268 of isolation fence 266 is typically included in the proximal open for leading to channel 264 274 and lead to isolation structure 272 distal openings 276 between the region that extends.The length L of bonding pad 268_conTwo can be more than The maximum penetration D of secondary flowing 282_p, in this case, secondary flow 282 may extend in bonding pad 268 without being weighed Newly it is directed to isolated area 270 (as shown in Figure 2 D).Alternatively, as shown in Figure 2 F, bonding pad 268, which can have, is less than maximum penetration depth Spend D_pLength, in this case, secondary flow 282 will extend through bonding pad 268 and draw again towards isolated area 270 It leads.In the latter case, the length L of bonding pad 268_c1And L_c2The sum of be more than maximum penetration D_pSo that secondary flow 282 It may not extend in isolated area 270.No matter the length L of bonding pad 268_conIt is greater than penetration depth D_pOr the length of bonding pad 268 Spend L_c1And L_c2The sum of be more than penetration depth D_p, do not exceed maximum speed V_maxFlowing of the first medium 254 in channel 264 278 will all generate with penetration depth D_pSecondary flow, and isolation fence 266 isolated area 270 in speck body (not Show, but can be identical or substantially similar as speck body 246 shown in Fig. 2 C) it will not be by the first medium in channel 264 254 flowing 278 is pulled out from isolated area 270.Flowing 278 in channel 264 will not be by the sundries from channel 264 (not Show) it brings into the isolated area 270 of isolation fence 266.In this way, diffusion be component in first medium 254 in channel 264 can To be moved to the exclusive mechanism in the second medium 258 in the isolated area 270 for completely cutting off fence 266 from channel 264.Similarly, expand The component dissipated in the second medium 258 in the isolated area 270 for being isolation fence 266 can be moved to channel 264 from isolated area 270 In first medium 254 exclusive mechanism.First medium 254 can be that medium identical with second medium 258 or first are situated between Matter 254 can be the medium different from second medium 258.Alternatively, first medium 254 and second medium 258 can start it is identical, Then become different, such as second medium is adjusted by one or more of isolated area 270 unit, or by changing Become the medium for flowing through channel 264.

As shown in Figure 2 E, the width W in the channel 264 in channel 264_ch(that is, transverse to 278 institute of arrow in Fig. 2 D is passed through The direction that is taken of direction of the fluid media (medium) flowing in the channel of instruction) it may be substantially perpendicular to the width W of proximal open 274_con1 And therefore it is basically parallel to the width W of distal openings 276_con2.However, the width W of proximal open 274_con1And distal openings 276 width W_con2It need not be substantially perpendicular to each other.For example, the width W of proximal open 274_con1The axis (not shown) oriented With the width W of distal openings 276_con2Angle between another axis oriented can not be vertical, therefore not be 90 °. The example of optional orientation angles includes the angle in following any range：About 30 ° to about 90 °, about 45 ° to about 90 °, about 60 ° extremely About 90 ° etc..

In the various embodiments of isolation fence (such as 124,126,128,130,224,226,228 or 266), isolated area (such as 240 or 270) are configured to contain multiple speck bodies.In other embodiments, isolated area, which can be configured as, only includes One, two, three, four, five or similar relatively few number of speck body.Therefore, the volume of isolated area can be For example, at least 1x10⁶、2x10⁶、4x10⁶、6x10⁶Cu μ m or more.

In the various embodiments of isolation fence, the width W in the channel (for example, 122) at proximal open (for example, 234)_ch It can be in any one of following range range：About 50-1000 microns, 50-500 microns, 50-400 microns, 50-300 it is micro- Rice, 50-250 microns, 50-200 microns, 50-150 microns, 50-100 microns, 70-500 microns, 70-400 microns, 70-300 it is micro- Rice, 70-250 microns, 70-200 microns, 70-150 microns, 90-400 microns, 90-300 microns, 90-250 microns, 90-200 it is micro- Rice, 90-150 microns, 100-300 microns, 100-250 microns, 100-200 microns, 100-150 microns and 100-120 microns. In some other embodiments, the width W in the channel (for example, 122) at proximal open (such as 234)_chIt can be in about 200-800 In the range of micron, 200-700 microns or 200-600 microns.It the above is only example, and the width W in channel 122_chIt can locate In (for example, the range limited by any endpoint listed above) within the scope of other.In addition, in channel in addition to completely cutting off fence In region except at proximal open, the W in channel 122_chIt can be selected as in any range in these ranges.

In some embodiments, isolation fence has about 30 to about 200 microns or about 50 to about 150 microns of height. In some embodiments, the sectional area for completely cutting off fence is about 1x10⁴–3x10⁶Square micron, 2x10⁴–2x10⁶Square micron, 4x10⁴– 1x10⁶Square micron, 2x10⁴–5x10⁵Square micron, 2x10⁴–1x10⁵Square micron or about 2x10⁵–2x10⁶Square micron. In some embodiments, bonding pad is with about 100 to about 500 microns, 200 to about 400 microns or about 200 to about 300 microns of section Face width.

In the various embodiments of isolation fence, the height H in the channel (for example, 122) at proximal open (for example, 234)_ch It can be in any one of following range range：20-100 microns, 20-90 microns, 20-80 microns, 20-70 microns, 20- 60 microns, 20-50 microns, 30-100 microns, 30-90 microns, 30-80 microns, 30-70 microns, 30-60 microns, 30-50 it is micro- Rice, 40-100 microns, 40-90 microns, 40-80 microns, 40-70 microns, 40-60 microns or 40-50 microns.It the above is only and show Example, and the height H of channel (such as 122)_chOther ranges be may be at (for example, being defined by any endpoint listed above Range) in.In the region in addition at the proximal open of isolation fence in channel, the height H in channel 122_chIt can be chosen It is selected as at any one within the scope of these.

In the various embodiments of isolation fence, the sectional area in the channel (for example, 122) at proximal open (for example, 234) It can be in any one of following range range：500-50,000 square microns, 500-40,000 square microns, 500- 30,000 square microns, 500-25,000 square microns, 500-20,000 square microns, 500-15,000 square microns, 500- 10,000 square microns, 500-7,500 square microns, 500-5,000 square microns, 1,000-25,000 square microns, 1, 000-20,000 square microns, 1,000-15,000 square microns, 1,000-10,000 square microns, 1,000-7,500 squares Micron, 1,000-5,000 square microns, 2,000-20,000 square microns, 2,000-15,000 square microns, 2,000-10, 000 square micron, 2,000-7,500 square microns, 2,000-6,000 square microns, 3,000-20,000 square microns, 3, 000-15,000 square microns, 3,000-10,000 square microns, 3,000-7,500 square microns or 3,000 to 6,000 squares Micron.Aforementioned is only example, and the section in the channel (for example, 122) at proximal open (for example, 234) may be at it In his range (for example, the range limited by any endpoint listed above).

In the various embodiments of isolation fence, the length L of bonding pad (for example, 236)_conIt may be in following range Either one or two of：About 1-600 microns, 5-550 microns, 10-500 microns, 15-400 microns, 20-300 microns, 20-500 microns, 40-400 microns, 60-300 microns, 80-200 microns or about 100-150 microns.It the above is only example, and bonding pad (such as 236) length L_conThe range different from aforementioned exemplary be may be at (for example, the model limited by any endpoint listed above Enclose) in.

In the various embodiments of isolation fence, the width of the bonding pad (for example, 236) at proximal open (for example, 234) Spend W_conIt may be at any of following range：20-500 microns, 20-400 microns, 20-300 microns, 20-200 microns, 20-150 microns, 20-100 microns, 20-80 microns, 20-60 microns, 30-400 microns, 30-300 microns, 30-200 microns, 30-150 microns, 30-100 microns, 30-80 microns, 30-60 microns, 40-300 microns, 40-200 microns, 40-150 microns, 40-100 microns, 40-80 microns, 40-60 microns, 50-250 microns, 50-200 microns, 50-150 microns, 50-100 microns, 50-80 microns, 60-200 microns, 60-150 microns, 60-100 microns, 60-80 microns, 70-150 microns, 70-100 microns and 80-100 microns.Aforementioned only example, and the width W of the bonding pad (for example, 236) at proximal open (for example, 234)_con It can be different from aforementioned exemplary (for example, the range limited by any endpoint listed above).

In the various embodiments of isolation fence, the width of the bonding pad (for example, 236) at proximal open (for example, 234) W_conIt can be at least with the isolation fence speck body to be used for (for example, it may be the life of T cell, B cell or ovum or embryo Object cell) full-size it is equally big.For example, drop by be placed into isolation fence proximal open 234 at bonding pad 236 width W_conIt may be at any one of following range range：About 100 microns, about 110 microns, about 120 microns, about 130 microns, about 140 microns, about 150 microns, about 160 microns, about 170 microns, about 180 microns, about 190 microns, it is about 200 micro- Rice, about 225 microns, about 250 microns, about 300 microns or about 100-400 microns, about 120 microns -350 microns, about 140-200- 200,300 microns or about 140-200 microns.Aforementioned only example, and the bonding pad at proximal open (for example, 234) The width W of (for example, 236)_conIt can be different from aforementioned exemplary (for example, the range limited by any endpoint listed above).

In the various embodiments of isolation fence, the width W of the proximal open of bonding pad_prCan at least with isolation fence institute The full-size of the speck body to be used for (for example, biological speck body of such as cell) is equally big.For example, width W_prCan be about 50 microns, about 60 microns, about 100 microns, about 200 microns, about 300 microns or can be in about 50-300 microns, about 50-200 Micron, about 50-100 microns, about 75-150 microns, about 75-100 microns or about 200-300 microns.

In the various embodiments of isolation fence, the length L of bonding pad (such as 236)_conWith the company at proximal open 234 Meet the width W in area (such as 236)_conRatio can be greater than or equal to following any ratio：0.5、1.0、1.5、2.0、2.5、 3.0,3.5,4.0,4.5,5.0,6.0,7.0,8.0,9.0,10.0 or bigger.Aforementioned is only example, and the length of bonding pad 236 Spend L_conWith the width W of the bonding pad 236 at proximal open 234_conRatio can be different from aforementioned exemplary.

In the various embodiments of microfluidic device 100,200,230,250,280,290,320,600,700, V_maxIt can set It is set to about 0.2,0.3,0.4,0.5,0.6,0.7,0.8,0.9,1.0,1.1,1.2,1.3,1.4 or 1.5 μ l/ seconds.

In the various embodiments of the microfluidic device with isolation fence, completely cut off the isolated area (for example, 240) of fence Volume can be for example, at least 5x10⁵、8x10⁵、1x10⁶、2x10⁶、4x10⁶、6x10⁶、8x10⁶、1x10⁷、5x10⁷、1x10⁸、 5x10⁸Or 8x10⁸Cu μ m or bigger.In the various embodiments of the microfluidic device with isolation fence, completely cut off fence Volume can be about 5x10⁵、6x10⁵、8x10⁵、1x10⁶、2x10⁶、4x10⁶、8x10⁶、1x10⁷、3x10⁷、5x10⁷Or about 8x10⁷ Cu μ m or bigger.In some other embodiments, the volume for completely cutting off fence can be about 1 nanoliter to about 50 nanoliters, 2 nanoliters To about 25 nanoliters, 2 nanoliters to about 20 nanoliters, about 2 nanoliters to about 15 nanoliters or about 2 nanoliters to about 10 nanoliters.

In various embodiments, microfluidic device has the isolation fence configured in any embodiment as discussed in this article, Wherein microfluidic device has about 5 to about 10 isolation fences, about 10 to about 50 isolation fences, about 100 to about 500 isolations Fence；About 200 to about 1000 isolation fences, about 500 to about 1500 isolation fences, about 1000 to about 2000 isolation fences Or about 1000 to about 3500 isolation fences.Isolation fence need not be all of the same size, and may include a variety of Configuration (for example, different in width, different characteristic in isolation fence).

In some other embodiments, there is microfluidic device the isolation configured in any embodiment as discussed in this article to enclose Column, wherein microfluidic device have about 1500 to about 3000 isolation fences, about 2000 to about 3500 isolation fences, about 2500 To about 4000 isolation fences, about 3000 to about 4500 isolation fences, about 3500 to about 5000 isolation fences, about 4000 to About 5500 isolation fences, about 4500 to about 6000 isolation fences, about 5000 to about 6500 isolation fences, about 5500 are to about 7000 isolation fences, about 6000 to about 7500 isolation fences, about 6500 to about 8000 isolation fences, about 7000 are to about 8500 isolation fences, about 7500 to about 9000 isolation fences, about 8000 to about 9500 isolation fences, about 8500 are to about 10,000 isolation fences, about 9000 to about 10,500 isolation fences, about 9500 to about 11000 isolation fences, about 10, 000 to about 11,500 isolation fence, about 10,500 to about 12,000 isolation fences, about 11,000 to about 12,500 isolations Fence, about 11,500 to about 13,000 isolation fences, about 12,000 to about 13,500 isolation fences, about 12,500 are to about 14,000 isolation fences, about 13,000 to about 14,500 isolation fences, about 13,500 to about 15,000 isolation fences, about 14,000 to about 15,500 isolation fences, about 14,500 to about 16,000 isolation fences, about 15,000 to about 16,500 Completely cut off fence, about 15,500 to about 17,000 isolation fences, about 16,000 to about 17,500 isolation fences, about 16,500 to About 18,000 isolation fences, about 17,000 to about 18,500 isolation fences, about 17,500 to about 19,000 isolation fences, About 18,000 to about 19,500 isolation fences, about 18,500 to about 20,000 isolation fences, about 19,000 to about 20,500 A isolation fence, about 19,500 to about 21,000 isolation fences or about 20,000 to about 21,500 isolation fences.

Fig. 2 G show the microfluidic device 280 according to one embodiment.Show that microfluidic device 280 is micro- in Fig. 2 G The stylized figure of fluid device 100.In fact, microfluidic device 280 and its composition loop element (such as channel 122 and isolation Fence 128) it will be with the size being discussed herein.The tool of microfluidic circuit 120 is there are two port 107 and with four shown in Fig. 2 G The flow region 106 in a difference channel 122.Microfluidic device 280 further comprises the multiple isolations opened from each channel 122 Fence.In the microfluidic device shown in Fig. 2 G, isolation fence has geometry similar with fence shown in fig. 2 C, therefore With both bonding pad and isolated area.Therefore, microfluidic circuit 120 include be swept region (for example, channel 122 and connection The maximum penetration D in secondary flow 244 in area 236_pInterior part) and the region that is not swept (for example, isolated area 240 With bonding pad 236 not in the maximum penetration D of secondary flow 244_pInterior part).

Fig. 3 A to Fig. 3 B show can be used for operating and observe microfluidic device according to the present invention (such as 100,200, 230,280,250,290,320) the various embodiments of system 150.As shown in Figure 3A, system 150 may include being configured as Accommodate the structure (" nest ") 300 of 100 (not shown) of microfluidic device or any other microfluidic device described herein.Nest 300 May include can with (for example, the photic dynamic electrical equipment 100) interface of microfluidic device 320 and provide from power supply 192 to miniflow The socket 302 of the electrical connection of body equipment 320.Nest 300 may further include integrated electric signal and generate subsystem 304.Telecommunications Number generate subsystem 304 can be configured as to socket 302 provide bias voltage so that when microfluidic device 320 is by socket 302 When holding, bias voltage is applied in a pair of electrodes in microfluidic device 320.Therefore, electric signal generation subsystem 304 can To be a part for power supply 192.The ability for applying bias voltage to microfluidic device 320 is not meant to work as microfluidic device 320 will apply bias voltage always when being kept by socket 302.It but in most cases, will be intermittently (for example, only needing When wanting) apply bias voltage, in order to the generation of the electric power (such as dielectrophoresis or electrowetting) in microfluidic device 320.

As shown in Figure 3A, nest 300 may include printed circuit-board assembly (PCBA) 322.Electric signal generates subsystem 304 can To be mounted on and be electrically integrated in PCBA 322.Example support part further includes the socket 302 being mounted on PCBA 322.

In general, it will include waveform generator (not shown) that electric signal, which generates subsystem 304,.Electric signal generates subsystem 304 It including oscillograph (not shown) and/or can also be configured as amplifying the waveform amplifying circuit of the waveform received from waveform generator (not shown).Oscillograph can be configured as measurement and is supplied to the microfluidic device kept by socket 302 (if present) 320 waveform.In certain embodiments, oscilloscope measurement in 320 nearside of microfluidic device (and in the remote of waveform generator Side) position at waveform, be actually applied to the waveform of equipment so that it is guaranteed that more accurately measuring.It is obtained from oscilloscope measurement Data can for example be provided as feedback to waveform generator, and waveform generator can be configured as based on this feedback To adjust its output.The example of one suitable combined waveform generator and oscillograph is Red Pitaya^TM。

In certain embodiments, nest 300 further includes controller 308, such as generates son for sensing and/or controlling electric signal The microprocessor of system 304.The example of suitable microprocessor includes Arduino^TMMicroprocessor, such as Arduino Nano^TM.Controller 308 can be used for executing function and analysis, or (can in figure 1A be shown) with Master controller 154 Communication is to execute function and analysis.In the embodiment as shown in fig. 3 a, controller 308 passes through interface 310 (such as plug or connection Device) it is communicated with master controller 154.

In some embodiments, nest 300 may include that electric signal generates subsystem 304 comprising Red Pitaya^TMWaveform Generator/oscillograph unit (" Red Pitaya units ") and the waveform that is generated by Red Pitaya units of amplification simultaneously transmit amplification Voltage to microfluidic device 100 waveform amplifying circuit.In some embodiments, Red Pitaya units are configured to measure Then amplification voltage at microfluidic device 320 adjusts the output voltage of their own so that microfluidic device 320 as needed The measurement voltage at place is desired value.In some embodiments, waveform amplifying circuit can have by one on PCBA 322 To+6.5V Zhi -6.5V the supplies of electric power that DC-DC converter generates, it is up to 13Vpp's to be generated at microfluidic device 100 Signal.

As shown in Figure 3A, support construction 300 may further include thermal control sub-system 306.Thermal control sub-system 306 can To be configured to adjust the temperature of the microfluidic device 320 kept by support construction 300.For example, thermal control sub-system 306 can be with Including Peltier (Peltier) thermal power unit (not shown) and cooling unit (not shown).Peltier thermoelectric equipment, which can have, matches It is set to the first surface at least one surface interfaces of microfluidic device 320.Cooling unit can be that for example cooling block is not ( Show), such as liquid cooling aluminium block.The second surface (for example, surface opposite with first surface) of Peltier thermoelectric equipment can To be configured as the surface interfaces with this cooling block.Cooling block may be coupled to fluid path 314, and fluid path 314 is matched Being set to makes cooling fluid cycle through cooling block.In the embodiment as shown in fig. 3 a, support construction 300 include entrance 316 and Cooling fluid is introduced fluid path 314 and led to by outlet 318 to receive the cooling fluid from external storage (not shown) Then cooling fluid is returned to external storage by supercooling block.In some embodiments, Peltier thermoelectric equipment, cooling list Member and/or fluid path 314 may be mounted on the shell 312 of support construction 300.In some embodiments, thermal control subsystem System 306 is configured as adjusting the temperature of Peltier thermoelectric equipment, to realize the target temperature of microfluidic device 320.Peltier The temperature adjusting of thermal power unit can for example pass through such as Pololu^TMThermoelectric power source (Pololu robot technology and electronics group) Thermoelectric power source realize.Thermal control sub-system 306 may include feedback circuit, the temperature value such as provided by analog circuit. Alternatively, feedback circuit can be provided by digital circuit.

In some embodiments, nest 300 may include the thermal control sub-system 306 for having feedback circuit, the feedback circuit It is analog voltage-dividing device circuit (not shown) comprising and resistor (such as resistance is 1kOhm+/- 0.1%, temperature coefficient is +/- 0.02ppm/C0) and NTC thermistor (such as nominal resistance be 1kOhm+/- 0.01%).In some cases, thermal control system System 306 measures the voltage for carrying out self-feedback ciucuit, then uses calculated temperature value as onboard pid control circuit algorithm Input.Output from pid control circuit algorithm can drive such as Pololu^TMOrientation on motor driver (not shown) and arteries and veins Bandwidth modulation signals pin is rushed, to activate thermoelectric power source, to control Peltier thermoelectric equipment.

Nest 300 may include serial port 324, allow the microprocessor of controller 308 via 310 (not shown) of interface It is communicated with Master controller 154.In addition, the microprocessor of controller 308 can generate subsystem 304 and thermal control with electric signal Subsystem 306 communicates (for example, via Plink tools (not shown)).Therefore, pass through controller 308, interface 310 and serial The combination of port 324, electric signal generate subsystem 304 and thermal control sub-system 306 and can be communicated with Master controller 154. In this way, master controller 154 can especially be calculated by executing the scaling adjusted for output voltage come auxiliary electric signal Generate subsystem 304.The graphic user interface (GUI) provided via the display equipment 170 for being coupled to Master controller 154 (not shown) can be configured as to draw generates the temperature that subsystem 304 obtains from thermal control sub-system 306 and electric signal respectively And Wave data.Alternatively or additionally, GUI can allow to update controller 308, thermal control sub-system 306 and electric signal Generate subsystem 304.

As described above, system 150 may include imaging device 194.In some embodiments, imaging device 194 includes light Mod subsystem 330 (referring to Fig. 3 B).Light modulating subsystem 330 may include digital mirror device (DMD) or miniature shutter array System (MSA), it is therein any one can be configured to receive the light from light source 332 and by the subset of the light received It is transferred in the optical system of microscope 350.Alternatively, light modulating subsystem 330 may include generating the light of their own (and therefore not Need light source 332) equipment, such as organic light emitting diode display (OLED), liquid crystal on silicon (LCOS) equipment, ferroelectric liquid crystals Silicon equipment (FLCOS) or transmissive type liquid crystal display (LCD).Light modulating subsystem 330 can be such as projecting apparatus.Therefore, light Mod subsystem 330 being capable of emitting structural light and unstructured light.Suitable light modulating subsystem 330 another example is come From Andor technologies^TMMosaic^TMSystem.In certain embodiments, the image-forming module 164 and/or power plant module of system 150 162 can control light modulating subsystem 330.

In certain embodiments, imaging device 194 further includes microscope 350.In such embodiments, nest 300 and light Mod subsystem 330 can be configured separately on microscope 350.Microscope 350 can be such as research on standard grade Light microscope or fluorescence microscope.Therefore, nest 300 may be configured on the objective table 344 mounted on microscope 350 and/ Or light modulating subsystem 330 may be configured on the port of microscope 350.In other embodiments, described herein Nest 300 and light modulating subsystem 330 can be microscope 350 global facility.

In certain embodiments, microscope 350 may further include one or more detectors 348.In some realities It applies in example, detector 348 is controlled by image-forming module 164.Detector 348 may include eyepiece, charge coupling device (CCD), phase Machine (such as digital camera) or any combination thereof.If there is at least two detectors 348, then a detector can be for example Quick frame rate camera, and another detector can be high-sensitivity camera.In addition, microscope 350 may include being configured to It receives the reflected light from microfluidic device 320 and/or transmitting light and gathers at least part of reflected light and/or transmitting light Optical system of the coke on one or more detectors 348.Microscopical optical system can also include for different detectors Different tube lens (not shown) so that the final magnification on each detector can be different.

In certain embodiments, imaging device 194 is configured at least two light sources.It is, for example, possible to use first Light source 332 generates structured light (for example, via light modulating subsystem 330), and can be provided using second light source 334 Unstructured light.First light source 332 can generate the structured light for photic electric excitation and/or fluorescence excitation, and second Light source 334 may be used to provide bright field illumination.In these embodiments, power plant module 164 can be used for controlling first light source 332 And image-forming module 164 can be used for controlling second light source 334.The optical system of microscope 350 can be configured as (1) and work as equipment When being kept by nest 300, receives the structured light from light modulating subsystem 330 and structured light is focused on into microfluidic device On at least first area in (such as photic electrical equipment), and (2) receive reflected light and/or hair from microfluidic device It penetrates light and focuses at least part of this reflected light and/or transmitting light on detector 348.Optical system can also by with It is set to when equipment is kept by nest 300 and receives the unstructured light from second light source and unstructured light is focused on into microfluid On at least second area of equipment.In certain embodiments, the first area of microfluidic device and second area can be overlappings Region.For example, first area can be the subset of second area.

In figure 3b, first light source 332 is illustrated as providing the light to light modulating subsystem 330, light modulating subsystem 330 to The optical system of the microscope 350 of 355 (not shown) of system provides structured light.Second light source 334 is illustrated as via beam splitter 336 Non-structured light is supplied to optical system.Structured light from light modulating subsystem 330 and from the non-of second light source 334 Structured light is advanced from beam splitter 336 across optical system together, the second beam splitter of arrival (or dichroic filter 338, it depends on The light provided by light modulating subsystem 330), wherein light is reflected to down reaches sample plane 342 by object lens 336.Then, come It is returned up from the reflected light and/or transmitting light of sample plane 342 across object lens 340, beam splitter and/or dichroic is passed through to filter Device 338, and reach dichroic filter 346.A part of light for only reaching dichroic filter 346 passes through and reaches inspection Survey device 348.

In some embodiments, second light source 334 emits blue light.It is flat from sample using dichroic filter 346 appropriate The blue light that face 342 is reflected can pass through dichroic filter 346 and reach detector 348.On the contrary, coming from light modulating subsystem 330 structured light is reflected from sample plane 342, but is not passed through dichroic filter 346.In this illustration, dichroic is filtered Light device 346 filters out the visible light that wavelength is more than 495nm.If the light emitted from light modulating subsystem does not include shorter than 495nm This filter out of any wavelength, the then light from light modulating subsystem 330 just calculates completion (as shown in the figure).In fact, if coming Light from light modulating subsystem 330 includes the shorter than wavelength of 495nm (for example, blue wavelength), then from light modulating subsystem Some light will pass through filter 346 to reach detector 348.In such embodiments, filter 346 is for changing from the first light Source 332 and second light source 334 reach the balance between the light quantity of detector 348.If first light source 332 is significantly stronger than the second light Source 334, then this can be beneficial.In other embodiments, second light source 334 can emit feux rouges, and dichroic filters Device 346 can filter out the visible light (such as visible light with the shorter than wavelength of 650nm) in addition to feux rouges.

Surface is modified.Surface for manipulating and storing the material of biomaterial, equipment and/or device can have non-needle To natural characteristic that is short-term and/or optimizing for a long time with material, this material can include but is not limited to speck body (including But be not limited to biological speck body, such as biological cell), biomolecule, the segment of biomolecule or biological speck body and they Any combinations.Modified material, one or more surfaces of device are one or more of biological to reduce and contact The associated undesirable phenomenon of one or more of self-faced of material may be useful.In other embodiments, increase The surface nature of strong material, equipment and/or device with may be by desired characteristic introducing surface it is useful, to expand material, Processing, manipulation or the working ability of equipment and/or device.For this reason, it may be necessary to can modified surface with reduce undesirable property or Introduce the molecule of desired property.

Compound for modified surface.In various embodiments, surface modifying compound may include that surface modification is matched Body, can be non-polymeric portions, such as moieties or substituted moieties, as fluoroalkyl moiety is (including but unlimited In perfluoroalkyl moieties), covalently it is modified the surface attached by it.Surface modifying compound further includes coupling part, the company Socket part point is the group that surface modified ligand is covalently attached to surface, as shown schematically in reaction equation 1.Covalent modified surface With the surface modified ligand being attached by the link group LG, the link group LG is coupling part and surface functional group (including hydrogen Oxide, oxide, amine or sulphur) reaction product.

Reaction equation 1.

In some embodiments, surface modifying compound may include forming straight chain (for example, at least ten carbon or at least 14, the straight chain of 16,18,20,22 or more carbon) carbon atom, and can be with right and wrong branch alkyl moieties.In some embodiments In, alkyl may include the alkyl (for example, some carbon in alkyl can be fluorinated or perfluorinated) of substitution.In some embodiments In, the alkyl may include the first segment for being connected to the second segment, and the first segment may include perfluoroalkyl, the second segment May include unsubstituted alkyl, wherein the first segment and the second segment can be with direct or indirect connections (for example, passing through ehter bond Mode).The first segment of alkyl can be located at the distal side of the link group, and the second segment of alkyl can be located at the close of coupling part Side.

In various embodiments, surface modifying compound can be with the structure of Formulas I：

Wherein coupling part V is-P (O) (OH) Q- or-Si (T)₂W；W is-T ,-SH or-NH₂, and the company of being configured as It is connected to the part on surface；Q is-OH and is arranged to be connected to the part on surface；And T is OH, OC_1-3Alkyl or Cl.R It is hydrogen or fluorine, M is hydrogen or fluorine.Each example of h is independently 2 or 3 integer；J is 0 or 1；K is 0 or 1；M is 0 or 1 to 25 Integer；And n is 0 or 1 to 25 integer.In some other embodiments, (n+ [(h+j) k]+m) and can be 11 To 25 integer.In some embodiments, M is hydrogen.In various embodiments, m is 2.In some embodiments, k is 0.At it In his embodiment, k is 1.In various embodiments, j is 1.For the compound of Formulas I, when k is integer 1, then m can be at least It is hydrogen for 2 and M.For compound of formula I, when k is 0 and R is fluorine, then it is hydrogen that m, which can be at least 2 and M,.

In various embodiments, when surface modifying compound has the structure of Formulas I, coupling part V can be-Si (T)₂W, wherein T and W are as defined above.W can be OC_1-3Alkyl or Cl.W can be methoxyl group, ethyoxyl or propoxyl group.At some In embodiment, W can be methoxyl group.T can be OC_1-3Alkyl or Cl.In various embodiments, coupling part V is-Si (OMe)₃.In various other embodiments, it is OH that V, which can be-P (O) (OH) Q, wherein Q,.

The surface modifying compound of formula 1 can have the preferred atomicity range for the linear backbone for constituting the compound.Such as Defined in upper, each segment of 1 compound of constitutional formula can have a certain range of size.Therefore, the compound of formula 1 can be with With repetitive unit as defined above so that (n+ [(h+j) k]+m) is equal to 25, will generate the total length of 26 atoms, wraps Include the end CR for being attached to coupling part₃Group.May include various in the case where (n+ [(h+j) k]+m) is equal to 25 Different compositions.For example, segment-[CR₂]_nThere can be n=23；–[(CH₂)_h-(O)_j]_kThere can be k=0；[CM₂]_mIt may There is m=2.Another example with same population (n+ [(h+j) k]+m) equal to 25 can be with segment-[CR₂]_n, wherein N=6；[(CH2) h- (O) j] k-wherein k=3, and include j=1 and h=2；And [CM₂]_mThere can be m=4.

In some embodiments, (n+ [(h+j) k]+m) and can be 11,13,15,17 or 21.In other embodiment In, (n+ [(h+j) k]+m) and can be 15 or 17.In other embodiments, (n+ [(h+j) k]+m) and can be 13 or 15.

In some embodiments, may exist one or more ehter bonds in compound of formula I.In some embodiments, j can To be 1.In some embodiments, wherein k and j, which is 1, m, can be at least 2.

In other embodiments, main chain carbon can be fluorinated.In some embodiments, main chain carbon can be fluoridized, Wherein CR₃And/or-[CR₂]_nAnd/or-[CM₂]_mEach R can be fluorinated.In some embodiments, one of compound Point can the other parts with fluorinated carbon backbone atom and compound can be with the carbon backbone atom being substituted with hydrogen.Example Such as, in some embodiments, CR₃And-[CR₂]_nSegment can have a non-master chain substituent of fluorine (for example, R be fluorine), and-[CM]_m– Segment can have the non-master chain substituent of hydrogen (such as M is hydrogen).In some embodiments, when R is fluorine, then k is 0.In other realities It applies in example, R can be fluorine and k be 1, j be 1 and h is 2.In various embodiments, M can be hydrogen.

In other embodiments, the compound of formula 1 can be hydrogenated by the silicon of alkene as described below to synthesize, and wherein m is extremely It is less hydrogen for 2, M.In some embodiments, it is hydrogen that m, which is 2 and M,.

Some in the various compounds of Formulas I can be more easily seen in the subgroup of compound described in following formula, But these formulas are never limited in the range of Formulas I.

In some embodiments, the compound of Formulas I may include the compound of formula 110：

CH₃(CH₂)_mSi(OC_1-3Alkyl)₃；

Formula 110

Wherein m is 9 to 23 integer.In some embodiments, m can be 11,13,15,17 or 19.In some other realities It applies in example, m can be 13 or 15.

In other embodiments, the compound of Formulas I may include the compound of formula 111：

CF₃(CF₂)_n(CH₂)₂Si(OC_1-3Alkyl)₃；

Formula 111

Wherein n can be 9 to 22 integer.Alternatively, n can be 11 to 17 integer.In some other embodiments, n Can be 9,11,13 or 15.In some embodiments, n can be 13 or 15.

In other embodiments, the compound of Formulas I may include the compound of formula 112：

CR₃(CR₂)_n(CH2)_hO(CH₂)_mSi(OC_1-3Alkyl)₃；

Formula 112

Wherein n is 3 to 19 integer；H is 2 or 3 integer；M is 2 to 18 integer.In some embodiments, R can be with It is fluorine.In some embodiments, n can be 3 to 11 integer, and h can be 2, and m can be 2 to 15 integer.

Alternatively, the compound of Formulas I may include the compound of formula 113：

CR₃(CR₂)_n(CM₂)_mP(O)(OH)₂；

Formula 113

Wherein n is 3 to 21 integer；M is 2 to 21 integer.In some embodiments of the compound of formula 113, R can be with It is fluorine.In some embodiments, M can be hydrogen.In various embodiments, n can be 5,7,9 or 11.In other embodiments, M can be 2,4,5,7,9,11 or 13.

For modified surface.It can be by the modified table of surface modifying compound as described herein (including compound of formula I) Face can be metal, metal oxide, glass or polymer.There can be some materials on the covalent modified surface introduced may include But it is not limited to silicon and its oxide, siloxanes, aluminium or its oxide (Al₂O₃), indium tantalum pentoxide (ITO), titanium dioxide (TiO₂), zirconium oxide (ZrO2), hafnium oxide (IV) (HfO₂), tantalum oxide (V) (Ta₂O₅) or any combination thereof.Surface can be this The chip or sheet material of a little materials, or can be in conjunction in device or equipment.In some embodiments, including in these materials The surface of any type can be bonded in microfluidic device as described herein.

Polymer may include any suitable polymer.Suitable polymer can include but is not limited to such as rubber, Plastics, elastomer, siloxanes, organosiloxane, such as dimethyl silicone polymer (" PDMS ") etc., can be ventilative.Its His example may include molding glass, patternable materials, such as siloxane polymer is (for example, photo-patterned polysiloxanes Or " PPS "), photoresist (for example, photoresist based on epoxy resin of SU8) etc..In other embodiments, The surface of material such as natural fiber or timber can pass through surface modifying compound as described herein (compound for including Formulas I) Functionalization is to introduce covalent modified surface.

Surface to be modified may include nucleophilic moiety, including but not limited to hydroxide, amino and mercaptan.On surface Nucleophilic moiety (for example, hydroxide (being known as oxide in some embodiments)) can be with surface modified compound as described herein Object (compound for including Formulas I) reacts, and is matched with being modified surface by siloxy the link group or phosphonate ester the link group Body is covalently attached onto surface, to provide functionalized surfaces.Surface to be modified may include natural nucleophilic moiety, or can be with It is handled with reagent (for example, Piranha solution) or by corona treatment to introduce nucleophilic moiety (for example, hydroxide (or referred to as oxide)).

In some embodiments, surface can individually or with any combinations be formed by any of above material.The surface can be with Including semiconductor substrate.In various embodiments, including the surface of semiconductor substrate may further include as described herein DEP or EW substrates.In some embodiments, the surface for including the semiconductor substrate with DEP or EW substrates can be as herein A part for the microfluidic device.

In some embodiments, modified surface can be at least one inward-facing of microfluidic device as described herein Surface.At least one surface can be a part for the flow region of microfluidic device (it may include channel), Huo Zheke To include the surface of such as enclosed construction of fence (may include isolation fence as described herein).

Covalent modified surface.Covalent modified surface may include surface modified ligand, can be non-polymer portion Point, such as moieties, substituted moieties, such as fluoroalkyl moiety (including but not limited to perfluoroalkyl moieties), and It can be any of the above described surface modified ligand, surface is covalently bound to by the link group, the link group is by connecting Part and surface part obtained by the reaction.The link group can be siloxy the link group or phosphonate ester the link group.

In some embodiments, surface modified ligand may include to be formed straight chain (for example, at least 10 carbon or at least 14, 16, the straight chain of 18,20,22 or more carbon) carbon atom, and can be with right and wrong branch alkyl moieties.In some embodiments, Alkyl may include the alkyl (for example, some carbon in alkyl can be fluorinated or perfluorinated) of substitution.In some embodiments, Alkyl may include the first segment for being bonded to the second segment, and the first segment may include perfluoroalkyl, and the second segment can wrap Unsubstituted alkyl is included, wherein the first segment and the second segment can be engaged directly or indirectly (for example, by way of ehter bond). First segment of alkyl can be located at the distal side of the link group, and the second segment of alkyl can be located at the close of the link group Side.

The covalent modified surface of Formula II.In some embodiments, covalent modified surface has the structure of Formula II：

Wherein it is surface；V is-P (O) (OY) W- or-Si (OZ)₂W.W is-O- ,-S- or-NH- and is connected to surface.Z is Key to the adjacent silicon atoms for being attached to surface or the key to surface.Y is the key to the adjacent phosphorus atoms for being attached to surface, or Person is the key to surface.For the covalent modified surface of Formula II, R, M, h, j, k, m and n are as defined above.When k is integer 1 When, then it is hydrogen that m, which is at least 2 and M,.When k is 0 and R is fluorine, then it is hydrogen that m, which is at least 2 and M,.The covalent modified surface of Formula II It can be described as the surface modified ligand being attached by the link group LG (such as Formula II A), wherein LG is linked to surface：

Covalent modified surface may include any surface of any combination of Formula II, such as change above for the surface of Formulas I Described in property compound.

In some embodiments, the covalent modified surface of Formula II can be the surface of formula 210：

WhereinIt is surface, and the oxygen of silicon atom attachment is also combined with the surface, and m is 11 to 23 integer.In some realities It applies in example, m can be 11,13,15,17 or 19.In some other embodiments, m can be 13 or 15.

In some other embodiments, the covalent modified surface of Formula II can be the surface of formula 211：

WhereinIt is surface, and the oxygen of silicon atom attachment is also combined with the surface, and n can be 9 to 22 integer.Alternatively, N can be 11 to 17 integer.In some other embodiments, n can be 7,9,11,13 or 15.In some embodiments, N can be 13 or 15.

In other embodiments, the covalent modified surface of Formula II can be the surface of formula 212：

WhereinIt is surface, and the oxygen of silicon atom attachment is also combined with the surface, and n is 3 to 21 integer, h is 2 or 3 Integer, and m is 2 to 21 integer.In some embodiments, R can be fluorine.In some embodiments, n can be 3 to 11 Integer, h can be 2, and m can be 2 to 15 integer.

Alternatively, the covalent modified surface of Formula II can be the surface of formula 213：

WhereinSurface, and the oxygen of phosphorus atoms attachment is also combined with the surface, n is 3 to 21 integer, m be 2 to 21 it is whole Number.In some embodiments of the compound of formula 113, R can be fluorine.In some embodiments, M can be hydrogen.In various realities It applies in example, n can be 5,7,9 or 11.In other embodiments, m can be 2,4,5,7,9,11 or 13.

In some embodiments, microfluidic device includes the flow region for being fluidly coupled to first entrance and first outlet, Flow region is configured to accommodate the flowing of first fluid medium.Microfluidic device may include leading to one of flow region or more Multiple chambers.Covalent modified surface can be the covalent modified substrate of microfluidic device and can flow region and/ Or below at least one chamber.In some embodiments, it is configured to the microfluidic device towards fluid completely or generally Whole inner surfaces have the covalent modified surface of Formula II.

Fig. 2 H depict the sectional view of the microfluidic device 290 comprising exemplary covalent modified surface 298.As shown, Covalent modified surface 298 (schematically showing) may include the lid 288 with the inner surface of substrate 286 294 and microfluidic device 290 The 292 intensive molecule of covalently bound single layer of inner surface.Covalent modified surface 298 can be disposed proximate to simultaneously inwardly miniflow On the essentially all inner surface 294,292 of the capsule 284 of body equipment 290, in some embodiments and as described above, include Surface (not shown) for the microfluidic circuit material for limiting loop element and/or structure in microfluidic device 290.For For in embodiment, covalent modified surface 298 can be provided only on one of microfluidic device 290 or some inner surfaces.

In the embodiment shown in Fig. 2 H, covalent modified surface 298 includes the single layer of alkyl-blocked siloxane molecule, Each molecule is via siloxy chain junctor 296 covalently bonded to the inner surface 292,294 of microfluidic device 290.For letter For the sake of list, the additional silicon oxide key for being linked to adjacent silicon atoms is shown, but the invention is not restricted to this.In some embodiments In, covalent modified surface 298 can be in its end towards capsule (i.e. in being not bonded to of the single layer of surface modified ligand 298 Surface 292,294 and part close to capsule 284) on include fluoroalkyl (such as fluorinated alkyl or perfluorinated alkyl).Although Fig. 2 H are discussed as the modified surface for having alkyl-blocked, but as described herein, and any suitable surface modified compound can be used Object.

Intrinsic surface.At least one surface of microfluidic device to be modified can be glass, metal, metal oxide or Polymer.It can combine in microfluidic device and can be modified with the covalent modified surface with the Formula II wherein introduced Some materials can include but is not limited to silicon and its oxide, siloxanes, aluminium or its oxide (Al₂O₃), indium tantalum oxide (ITO), titanium dioxide (TiO₂), zirconium oxide (ZrO2), hafnium oxide (IV) (HfO₂), tantalum oxide (V) (Ta₂O₅) or its any group It closes.Polymer may include any suitable polymer.Suitable polymer can include but is not limited to (such as rubber, plastics, Elastomer, siloxanes, organosiloxane, such as dimethyl silicone polymer (" PDMS ") etc.), can be ventilative.Other show Example may include molding glass, such as patternable of siloxane polymer (such as photo-patterned polysiloxanes or " PPS ") Material, photoresist (for example, photoresist based on epoxy resin of SU8) etc..

The physics and performance characteristics on covalent modified surface.In some embodiments, covalent modified surface can have and increase The hydrophobicity added.The increased hydrophobicity of modified surface can prevent biomaterial dirt.As used herein, surface smut refers to It is deposited on the amount of the material on microfluidic device surface indiscriminately, may include biomaterial such as protein and degradation The permanent or semipermanent deposition of product, nucleic acid and respective catabolite.This fouling can increase biological speck body to surface Adhesive capacity.In other embodiments, the increased hydrophobic property on covalent modified surface can reduce biological speck body on the surface Adherency, and it is unrelated with the adherency caused by surface smut.

The modification on surface can increase the durability, functionality and/or biocompatibility on surface.It is each in these characteristics It is a to be further conducive to survival ability (including growth rate and/or cell doubling rates), as described herein covalent modified The property or speck body or biomolecule of the bacterium colony formed on surface (including the surface with Formula II structure) are in modified surface Portability (feasibility for including outlet) in upper and the equipment with covalent modified surface and/or device.

In some embodiments, (it can be any surface as described herein, include the table of Formula II on covalent modified surface Face) there can be the thickness (for example, being less than about 7nm, being less than about 5nm or about 1.5 to 3.0nm) for being less than 10nm.This can change Property surface on advantageous thin layer is provided, especially with other hydrophobic materials of the typical thickness that forms about 30 to 50nm (such as The perfluor-tetrahydrofuran polymer of spin coating) compare.Data shown in table 1 are for as shown in Table through processing With the silicon/oxidative silicon surface with covalent modified surface.Contact-angle measurement value is obtained using static drop method.(Drelich, J.Colloid Interface Sci.179,37-50,1996.) thickness measured by ellipsometry.

Contact angle hysteresis measurement is carried out using Biolin Scientific contact angle angular instruments.By the OEW tables of chemical modification Face is placed in the pond of 5cSt silicone oil, which is mounted in transparent vessel.Then phosphate buffered saline (PBS) (PBS) liquid droplet distribution is arrived On surface in oil.Platinum (Pt) line electrode is inserted into drop, fixed water contact angle is measured.Next, in OEW substrates and inserting Enter with 30kHz frequencies to apply between the Pt conducting wires in PBS drops the AC voltages 10 seconds of 50Vppk.Then, the voltage of application is removed, Contact angle is measured again.Apply 50Vppk AC electricity by being subtracted from the original contact angle before applying voltage under zero-bias Contact angle after pressure under zero-bias calculates contact angle hysteresis.

Table 1. selectes the physical data on surface

T and Q are as described above.

The contact angle for being less than 10 degree of water on the contact angle for the modified surface observed and the silicon face of plasma cleaning On the contrary.Each in these surfaces is smaller than the wettable on natural silicon/oxidative silicon surface.

It may include infrared spectrum and/or x-ray photoelectron spectroscopy to be suitble to other analysis methods on characterization surface.

Another ideal characterisitics of modified surface of the present invention is a lack of autofluorescence, may depend on surface modifying compound Chemical property.

In some embodiments, covalent modified surface (surface for including Formula II) as described herein can form single layer. The homogeneity and uniformity on monolayer-modified surface can provide advantageous performance, especially have other work(on monolayer-modified surface In the case of energy attribute.For example, covalent modified surface (surface for including Formula II) as described herein can also include electrode active Change substrate, and optionally can also include dielectric layer, such as can with dielectrophoresis configuration or electrowetting configuration material, set It is found in standby and/or device.Compared with the single layer containing such as alkene or aromatic fractions, the perfluoroalkyl moieties of modified surface Degree of unsaturation shortage can make " charge traps " minimize.In addition, being formed in surface (including Formula II surface) described herein The dense accumulation property of single layer cation can be made to be driven through single layer and reach following metal, metal oxide, glass Or the possibility of polymer substrate minimizes.It is without being bound by theory, destroy lining by cation is added to substrate compositions Bottom surface may destroy the electrical properties of substrate, to reduce the ability of its electric functions.

In addition, the ability for introducing modified surface by covalent linkage can increase dielectric strength and the protection of modified surface Following material is not breakdown when applying electric field.Material with covalent modified surface (including Formula II surface) described herein, The uniformity and thinness of the dielectrophoresis or electrowetting surface of equipment and/or device are that optics causes in the material, equipment and/or device The advantageous benefit on this modified dielectrophoresis and/or electrowetting surface can be further provided in the case of dynamic.

The preparation method on covalent modified surface.May be used as the surface of the material of the component of device can assemble It is modified before the device.Alternatively, the device that can be constructed with modifying moieties or completely so that include by contact It all surface of the biomaterial of biomolecule and/or speck body (it may include biological speck body) while being modified.At some In embodiment, though there are different materials at different surfaces in equipment and/or device, the equipment and/or device it is whole A inside can also be modified.In some embodiments, the equipment and/or device partially or completely constructed can be such as this paper institutes The microfluidic device stated or its component.

Surface to be modified can be cleaned before modification can freely be used for the nucleophilic moiety ensured on surface Reaction, such as not by oil or adhesive coverage.Cleaning can be completed by any suitable method, including with including alcohol or third The solvent processing of ketone, supersound process, cleaning steam etc..It in some embodiments, will be at the oxygen plasma of surface to be modified Reason is handled, and the oxygen plasma treatment removes pollutant, can introduce on the surface simultaneously additional oxide (such as Hydroxide) part.This can advantageously provide more sites for being modified on the surface, to provide closer accumulation Modified surface layer.

In some embodiments, the method on covalent modified surface includes the following steps：The compound of surface and Formulas I is set to connect It touches：

Wherein V is-P (O) (OH) Q or-Si (T)₂W.W is-T ,-SH or-NH₂, and be configured as being connected to surface Part.Alternatively, when V is-P (O) (OH) Q, Q is-OH and is configured as being connected to the part on surface.T is OH, OC_1-3Alkane Base or Cl.R, each in M, h, j, k, m and n is as above for defined in compound of formula I.(n+[(h+j)·k]+m) Summation be 11 to 25 integer.In various embodiments, when k is integer 1, then it is hydrogen that m, which is at least 2 and M,；When k is 0 and R For fluorine when, then m be at least 2 and M be hydrogen.Compound of formula I is reacted with the nucleophilic moiety on surface；And form covalent modified surface. Any combinations or sub-combination of compound of formula I can be used, as described above.

In the various embodiments of this method, the covalent modified surface being thusly-formed can be single layer.

In some embodiments of this method, the compound of Formulas I can be the compound of formula 110：

CH₃(CH₂)_mSi(OC_1-3Alkyl)₃；

Formula 110

In the other embodiment of this method, the compound of Formulas I can be the compound of formula 111：

CF₃(CF₂)_n(CH₂)₂Si(OC_1-3Alkyl)₃；

Formula 111

Wherein n is 9 to 22 integer.Alternatively, n can be 11 to 17 integer.In other embodiments, n can be 11 To 17 integer.In some other embodiments, n can be 9,11,13 or 15.In some embodiments, n can be 13 or 15。

In the other embodiment of this method, the compound of Formulas I can be the compound of formula 112：

CR₃(CR₂)_n(CH2)_hO(CH₂)_mSi(OC_1-3Alkyl)₃；

Formula 112

Wherein n is 3 to 21 integer；H is 2 or 3 integer；M is 2 to 21 integer.In some embodiments, R can be with It is fluorine.In some embodiments, n can be 3 to 11 integer, and h can be 2, and m can be 2 to 15 integer.

Alternatively, surface can be contacted with the compound of the Formulas I for the compound that can be formula 113：

CR₃(CR₂)_n(CM₂)_mP(O)(OH)₂；

Formula 113

Contact procedure can be carried out by making surface be contacted with the liquid solution of the compound containing Formulas I.For example, surface The solution of the compound of the Formulas I containing 0.01mM, 0.1mM, 0.5mM, 1mM, 10mM or 100mM can be exposed to.The reaction can To carry out and can carry out at ambient temperature about 2 hours, 4 hours, 8 hours, 12 hours, 18 hours, 24 hours or therebetween Any value a period of time.The example of solvent includes but not limited to：Toluene, 1,3 pairs of trifluoro-benzenes or Fluorinert^TM(3M) fluorine Change solvent.If it does, acid such as acetic acid can be added in solution, to be improved by promoting the hydrolysis of tri-alkoxy group Reaction rate.

Alternatively, surface can be contacted with the gas phase of the compound containing Formulas I.In some embodiments, when by making surface When contacting progress reaction step in the gas phase with the compound of Formulas I, there is also the vapor of controlled quentity controlled variable.The vapor of controlled quentity controlled variable can With by being placed on the magnesium sulfate 7 hydrate of pre-selected amount in chamber identical with having the object on surface to be modified or capsule To provide.In other embodiments, it can be supplied by external vapor and the water of controlled quentity controlled variable is introduced into reative cell or capsule.Instead It should can be carried out under the decompression relative to atmospheric pressure.In some embodiments, decompression can be 100 supports or smaller.At other In embodiment, decompression can be less than 10 supports or be less than 1 support.

The reaction can carry out at a temperature of about 150 DEG C to about 200 DEG C.In various embodiments, reaction can be about It is carried out at a temperature of 150 DEG C, 155 DEG C, 160 DEG C, 165 DEG C, 170 DEG C, 175 DEG C, 180 DEG C, 185 DEG C or about 190 DEG C.Reaction can be with Be allowed to continue about 2h, 6h, 8h, 18h, for 24 hours, 48h, 72h, 84h or longer time.

In some embodiments, covalent modified surface can be with the structure of Formula II：

Wherein R, M, n, h, j, k, m and V are as described above, with any combinations.In some embodiments of this method, covalently change Property surface can with formula 210,211,212 or 213 as described above formula, have for each formula permissible element appoint What is combined.

In the various embodiments of this method, surface may include selected from the group being made of hydroxide, amino and mercaptan The nucleophilic moiety of group.Surface can be metal, metal oxide, glass, polymer.Metal surface can wrap Include silicon, silica, hafnium oxide, indium oxide tantalum, aluminium oxide.

In the various embodiments of this method, wherein the step of forming covalent modified surface can serve as a contrast in DEP substrates or EW It is carried out on bottom.The step of forming covalent modified surface may include at least one table in the microfluidic circuit element of microfluidic device Covalent modified surface is formed on face.Microfluidic circuit element may include wall, flow region, fence and electrode activation substrate, packet Include DEP or EW substrates.Surface in microfluidic circuit that can be covalent modified can be the fluid bearings towards microfluidic device Partial completely or generally whole surfaces.For example, in microfluidic device 200,230, it is all towards microfluidic channel 122 And the inner surface of top electrodes 210, the upper surface of electrode activation substrate 206, the microfluidic circuit material of fence 244,246,248 The surface (referring to Figure 1B, 1C, 2A, 2B) of material 116 can be modified.Similarly, in Fig. 2 D-2F, microfluidic circuit material 260 Inner surface, limit isolation fence 266 isolation structure 272 surface or all surface towards microfluidic circuit 262 can be with It is covalent modified by method described herein.

Unmixability medium.The movement of aqueous solution drop on the surface of a substrate can in region be distributed in one or more In flow region (it may include flow channel) and in the chamber (if present) for being fluidly connected to flow region It is carried out in interior water unmixability fluid media (medium).Water unmixability fluid media (medium) can have the kinematic viscosity more than pure water drop. Water unmixability fluid media (medium) can have the kinematic viscosity within the scope of about 1 centistoke (cSt) to about 15cSt, wherein 1cSt etc. In 1 millipascal or 1 centipoise (CPS).In some embodiments, water unmixability fluid media (medium) can have about 3cSt to about 10cSt or Viscosity within the scope of about 3cSt to about 8cSt.Water unmixability fluid media (medium) can be non-combustible at a temperature of at least 100 DEG C 's.In period in the aqueous drop that biological cell is processed, cultivated or is stored in water unmixability fluid media (medium), water Unmixability fluid media (medium) can be nontoxic to biological cell living.

Water unmixability fluid media (medium) can have low or very small solubility in water.Water unmixability fluid is situated between Matter can be dissolved when contacting (such as being separated with water) with water layer about 5% less than its total water capacity, 4%, 3%, 2%, 1% or Less than 1%.At a temperature of in the range of about 25 DEG C to about 38 DEG C, water unmixability fluid media (medium) can not dissolved not to be mixed more than water The volume of aqueous drop present in dissolubility fluid media (medium) about 5%, about 10%, about 15%, about 20%, about 25% or about 30%.In some embodiments, the dissolving of water unmixability fluid media (medium) is less than existing aqueous in water unmixability fluid media (medium) About the 20% of the volume of drop.

Water unmixability fluid media (medium) may include at least one with the main chain knot comprising the atom selected from carbon, silicon and oxygen The organic or organo-silicon compound of structure.In some embodiments, water unmixability fluid media (medium) may include it is more than one it is organic/ Organo-silicon compound, wherein more than one compound is polymer organic/organo-silicon compound, with polymer compound The molecular weight ranges of subunit.For example, polymer organic/organo-silicon compound can have two different Asias for constituting polymer Unit (such as copolymer), and each in two different subunits may be present in the repetition range with general formula AaBb Interior, wherein A and B are two different polymeric subunits, and a and b are the quantity that each subunit repeats.The quantity a and b repeated Individual integer is can not be, and can be the range of recurring unit.

In other embodiments, including the water unmixability fluid media (medium) of more than one organic/organo-silicon compound can be with The mixture of mixture, organo-silicon compound including organic compound or any combination thereof.Water unmixability fluid media (medium) can To include with any suitable mixing that will provide the different chemical structures of suitable performance and/or the compound of molecular weight Object.

The compound of water unmixability fluid media (medium), which can have, is less than about 1000Da, about 700Da, about 500Da or about 350Da Molecular weight.In other embodiments, the compound of water unmixability medium can be with than about 1000Da highers and still The molecular weight of suitable performance is provided.

In various embodiments, organic/organo-silicon compound of water unmixability fluid media (medium) can have backbone structure, It is carbon, silicon or oxygen wherein to constitute the atom of main chain.The substituent group of main chain carbon can be hydrogen or fluorine.In some embodiments, water is not Compatibility fluid media (medium) may include one or more of organo-silicon compound, and the main chain of wherein organo-silicon compound may include Silicon and oxygen atom.The silicon atom of organo-silicon compound can have carbon substituent group, can have hydrogen or fluoro substituents again.One In a little embodiments, the carbon substituent group of organo-silicon compound can be fluorine (such as fluoridized) entirely.In other embodiments, organic The carbon substituent group of silicon compound can be partially fluorinated.In various embodiments, the substitution of the carbon atom of organo-silicon compound Base can be no more than about 90% fluorine, 80% fluorine, 70% fluorine, 60% fluorine, 50% fluorine, 40% fluorine, 30% fluorine, 20% fluorine or less.

In other embodiments, the organic compound of water unmixability fluid media (medium) can have backbone structure, wherein structure Atom at main chain is carbon or oxygen.In some embodiments, the substituent group of main chain carbon can be hydrogen or fluorine.In other embodiment In, the substituent group of main chain carbon may include oxygen-containing part, such as ether, carbonyl or carbonate component.In some embodiments, water is not The organic compound of compatibility fluid media (medium) can have full carbon backbone chain structure.In some embodiments, water unmixability fluid The full carbon backbone chain structure of the organic compound of medium can have whole fluoro substituents (such as perfluorinated) on carbon atom.At it In his embodiment, the substituent group of organic compound can be partially fluorinated (such as not being fluoridized).In various embodiments In, include the carbon atom of the organic compound of the compound with full carbon backbone chain substituent group can be no more than about 90% fluorine, 80% fluorine, 70% fluorine, 60% fluorine, 50% fluorine, 40% fluorine or less.In some embodiments, water unmixability fluid media (medium) Suitable organic compound may include or can be single fluorine-substituted hydrocarbon, such as n octyl fluoride, 1- fluorine decane, 1- fluorine Dodecane or the 1- fluorine tetradecanes.

In other embodiments, the organic compound of water unmixability fluid media (medium) can not have fluorine on carbon and replace Base, but can have hydrogen substituent group.In some embodiments, the organic compound of water unmixability fluid media (medium) can have insatiable hunger And the ethylenic group in carbon-carbon bond, such as main chain carbon or at terminal position.

In some embodiments, selection will include considering including the appropriate compound in water unmixability fluid media (medium) Other properties of compound.In various embodiments, the compound used in water unmixability fluid media (medium) is suitble to be swashed Light, will not autofluorescence when projecting structured light in microfluidic device or daylight/laboratory illumination irradiation.

In other embodiments, the property of covalent modified hydrophobic surface can influence in water unmixability fluid media (medium) The selection of the suitable compound used.For example, covalent modified surface can have enough hydrophobicitys so that perfluorinated water is not Water droplet in compatibility fluid media (medium) can show sufficiently high surface tension so that water droplet cannot use as described herein Photoelectricity wetting configuration movement.

In some other embodiments, the property of microfluidic circuit material can influence to make in water unmixability fluid media (medium) The selection of the suitable compound.Return path materials expansion is set to may remain in acceptable limit by water unmixability fluid media (medium) In degree.For example, in some embodiments, if microfluidic circuit material includes having for the aryl substitution of SU8 or light patternable Organic siloxane, then can select include ring-type, the linear hydrocarbon of aryl or heteroaryl group, linear fluorocarbon or Carbon backbone chain compound is for using.

In other embodiments, microfluidic circuit material may include other materials, for example, without aryl substitution can light Cause patterned organosiloxane, and by using different compounds in water unmixability fluid media (medium), it can will be swollen Swollen limitation is within the acceptable limits.For example, compared with being exposed to water unmixability fluid media (medium) in advance, be less than about 40%, 30%, 20% or 10% expansion can be acceptable.However, in some embodiments, still can select not mix in water Cause the compound of expansion in dissolubility fluid media (medium).

In some embodiments, the compound of water unmixability fluid media (medium) can have carbon containing or oxygen atom main chain Organic compound.In some embodiments, organic compound can have the main chain containing carbon atom and without oxygen atom, and And further wherein atoms backbone is branch.In various embodiments, the organic compound of water unmixability fluid media (medium) The atoms backbone of branch be acyclic.The organic compound of the water unmixability fluid media (medium) of carbon backbone chain with branch can To be further free of any cyclisation part.

Although above-mentioned selection criteria can be used for selecting in one or more of water unmixability fluid media (medium)s to be included in Compound, and the compound that acceptable performance cannot be provided is eliminated, but acceptable water unmixability fluid media (medium) can be Multicomponent mixture, and may include individual organic or organo-silicon compound certain parts, when as water unmixability When the sole component of fluid media (medium), acceptable performance will not be provided.For example, when used alone, component may excessively high fluorine Change or may unacceptably expand microfluidic circuit material, but can or organo-silicon compound organic with other be applied in combination with Form water unmixability fluid media (medium).

Some the suitable organic compounds being used in water unmixability fluid media (medium) to independent or any category combinations can To include：Isohexadecane, ten difluoro hexane (HFE-7500,3MTM, NovecTM) of 2- (trifluoromethyl) -3- ethyoxyls, seven methyl Nonane (HMN), two (2- ethylhexyls) carbonic esters (TEGOSOFT DEC, (Evonik)) and (ten three fluoro- 1,1,2,2- tetrahydrochysene is pungent Base) tetramethyl disiloxane (Gelest, Cat#SIB 1816.0) or silicone oil (5 centistoke viscosity, Gelest Cat#DMS-T05).

Aqueous drop.Aqueous drop can contain one or more speck bodies, may include biological cell or bead.It is aqueous Drop contains the biologic that may include nucleic acid or protein.In some other embodiments, aqueous drop, which can contain, to be useful for The reagent of measurement can be any kind of reagent, such as the probe or chemical reagent of enzyme, antibody, fluorescent marker.

In some embodiments, aqueous drop can also include surfactant.Surfactant, which can increase water, not to be mixed The portability of aqueous drop in dissolubility fluid media (medium).In some embodiments, suitable surfactant may include it is non-from Sub- surfactant.In various embodiments, surfactant can be but not limited toBlock oxyalkylene is copolymerized Object, including F68 (ThermoFisher Cat.#2400032)；Aliphatic ester ethoxylated sorbitan alcohol, such as(Signa Aldrich Cat.#PI 379) or(Sigma Aldrich P1629)； 2,4,7,9 tetramethyl -5- decine -4,7- diol ethoxylates (TET, Sigma Aldrich Cat#9014-85-1)；Ethoxy Base nonionic fluorosurfactant, such asFS-30 (DuPont TM, Synquest Laboratories Cat.#2108-3-38).In some embodiments, lauryl sodium sulfate (SDS) may be used as surface work Property agent.In various embodiments, phosphate buffered saline (PBS) (PBS) may be used as surfactant.It can be by surfactant with about 1%, 3%, 5%, 10%, 15%, 20%, the range of about 25%v/v or any value therebetween is added in aqueous drop.

System.The present invention provides be used for transmission speck body that is compatible with aqueous medium and/or dissolving in aqueous medium, life The system of produce product and/or reagent.The system may include any microfluidic device for example disclosed herein (for example, with packet The microfluidic device of capsule containing base portion and microfluidic circuit structure, the wherein base portion include at least one with the upper surface of base portion The hydrophobic monolayer that some covalent combines).In addition, the system includes fluid media (medium) and aqueous drop, wherein fluid media (medium) and aqueous Drop is unmixability fluid.Fluid media (medium) can be any unmixability medium as described herein, and aqueous drop can be with Including any biomaterial as described herein and/or chemical reagent are (for example, protein, nucleic acid, detergent, surfactant Deng).

Kit.The present invention also provides suitable for transmitting speck that is compatible with aqueous medium and/or dissolving in aqueous medium The kit of body, biological product and/or reagent.Kit can include any microfluidic device disclosed herein (for example, miniflow Body equipment has the capsule comprising base portion and microfluidic circuit structure, and wherein base portion includes at least one with the upper surface of base portion Divide covalently bound hydrophobic monolayer).Kit, which can further include the fluid media (medium) unmixing with aqueous medium and other, to be had Reagent (such as surfactant etc.).

The method for manufacturing microfluidic device.The microfluidic device (such as device 400) of the present invention can pass through following steps Manufacture：(i) spacer element 108 is attached to the inner surface 428 of lid 110, which has at least one electrode, the electrode quilt It is configured for connection to AC voltage sources (not shown) (ii) and spacer element 108 (and associated lid 110) is attached to substrate 104 Dielectric surface 414, the substrate 104 have at least one electrode 418 for being configured to connect to AC voltage source (not shown), thus Spacer element 108 becomes between the inner surface 428 for being clipped in lid 110 and the dielectric surface 414 of substrate 104, middle cover 110 and substrate 104 are substantially parallel to each other orientation, and substrate 104, spacer element 108 and lid 110 collectively define and be configured to accommodate liquid Capsule 435, and (iii) pass through vapor deposition and form outer hydrophobic layer at least part of the inner surface 428 of lid 110 412, and outer hydrophobic layer 412 is formed at least part of the inner-dielectric-ayer of substrate 104 414.

By the vapor deposition of amphiphile, amphiphilic molecule, the single layer of dense packing, wherein amphiphilic may be implemented in hydrophobic layer 422 and 412 Molecule is covalently bound to the molecule of the interior dielectric surface 414 of base portion 104 and the inner surface 428 of lid 110 respectively.As described herein What self-association molecule and its equivalent can be vapor-deposited on the inner surface of microfluidic device.In order to obtain preferably accumulate it is close Degree, it includes such as alkyl that can be vapor-deposited at a temperature of at least 110 DEG C (for example, at least 120,130,140,150,160 etc.) The self-association molecule of the siloxanes of sealing end continue at least 15 hours (for example, at least 20,25,30,35,40,45 or more hours) Period.This vapor deposition is usually under vacuum and in water source such as epsom salt (i.e. MgSO₄·7H₂O in the presence of) It carries out.Typically, increase the improved characteristic of the temperature and duration generation hydrophobic layer 422 and 412 of vapor deposition.For example, By precleaning lid 110 (having spacer element 108) and substrate 104, vapor deposition processes can be improved.For example, this Precleaning may include solvent bath, such as acetone bath, ethanol bath or combinations thereof.Solvent bath may include Sonication.It can replace For ground or additionally, this precleaning may include that lid 110 (having spacer element 108) is handled in oxygen plasma cleaner With substrate 104.Oxygen plasma cleaner can for example be worked 60 seconds with 100W under vacuum.

Fig. 6 shows that the example of microfluidic device 600, microfluidic device 600 include with microfluidic channel 612,614 and The capsule of multiple chambers 616 and for capsule provide fluid drop 620 droplet generator 606.614 quilt of microfluidic channel It is configured to keep first fluid medium 624.Typically, first fluid medium is hydrophobic fluid, such as oil (such as silicone oil or fluorination Oil).Microfluidic channel 614 is connected to droplet generator 606 via interface 608, and interface 608 allows channel 614 to receive by drop The drop 620 that generator 606 generates.The drop 620 of reception is included in liquid unmixing in first fluid medium 624.In general, The drop received will include aqueous medium, can contain speck body, such as cell or bead or the examination for dissolving in aqueous medium Agent.Microfluidic channel 614 is also connected to each in multiple rooms 616, promotes received drop 620 (and from can not mix Close the drop 632 extracted out in the reservoir of the fluid in first fluid medium 624) enter in chamber 616 and in chamber 616 Between move.

The microfluidic channel 612 of device 600 is connected to the subset of chamber 616, and therefore via between these chambers 616 in succession It is connected to microfluidic channel 614.As shown, microfluidic channel 612 and chamber connected to it 616 are included in first fluid medium Unmixing fluid media (medium) 622 in 624.Thus, for example, fluid media (medium) 622 can be aqueous medium, such as cell culture medium. When fluid media (medium) 622 is cell culture medium, the chamber 616 containing culture medium may be used as the culture chamber of growth cell, and And microfluidic channel 612 can be to provide the perfusion channel of fresh cultured base flow.As discussed herein, fresh in perfusion channel The flowing of culture medium can be spread by the molecule between perfusion channel and culture chamber to be provided nutrients to chamber and is moved from chamber Except waste, to promote cell continued growth.

Fig. 7 shows another example of microfluidic device 700 comprising has microfluidic channel more than 612,614, first The capsule of chamber 716 and more than second a chambers 616 and droplet generator 606 for providing from fluid drop 620 to capsule.Figure 7 show that the modification of microfluidic device 600 shown in Fig. 6, middle chamber 616 are included in first fluid medium 624 and (are located at micro- In fluid channel 614) in unmixing medium 622, and chamber 616 is positioned directly in corresponding cavity across microfluidic channel 614 716 opposite of room.It should be configured with and be conducive to fluid drop 632 (including optionally speck body 630 or biomaterial) from selection chamber 616 Be moved to corresponding chamber 716, corresponding chamber 716 can be with treatment fluid drop (and any speck body 630 or biological material Material).

Another example of microfluidic device includes having more than 612,614, first a chamber 716 of microfluidic channel and more than second The capsule of a chamber 616 and for capsule provide fluid drop 620 droplet generator 606.The embodiment is presented in Fig. 7 Shown in microfluidic device 700 modification, middle chamber 616 is tapered at one end, to be tilted to chamber in microfluidic device It is moved convenient for microparticle when 616 tapered ends have lower potential energy (in applicable gravitational field) relative to non-tapered end To the interface of first fluid medium 624 and second fluid medium 622.

The microfluidic circuit formed by microfluidic channel 612,614 and chamber 616,716 is only example, and the present invention Cover many other configurations of channel and chamber.For example, in each in device 600 and 700,612 He of microfluidic channel The chamber 616 for being directly connected to channel 612 is optional feature.Therefore, device 600 and 700 can lack perfusion channel and culture Chamber.

In there are the embodiment of microfluidic channel 612, help limits channel 612 and/or the chamber being directly connected to 616 The substrate of (for example, base portion by forming channel and/or chamber) can have electrowetting configuration.Alternatively, however, it helps Electrowetting configuration can be lacked (for example, and instead can be with by limiting the substrate of channel 612 and/or the chamber 616 being directly connected to It is configured with DEP, or had not both had electrowetting and configured or configured without DEP).There is microfluidic channel 612 wherein simultaneously And the substrate for limiting channel 612 and/or the chamber being directly connected to 616 is helped to have in the embodiment of electrowetting configuration, substrate The outer hydrophobic surface that outer hydrophobic surface can be patterned as the substrate than helping to limit channel 614 is more hydrophilic.For example, institute as above It states, increased hydrophily may be implemented.

Droplet generator 606 and its provide drop any microfluidic circuit can be microfluidic device a part (or Person's integral part is either connected thereto) it can be as any microfluidic device as shown in the figure or described herein.Although scheming A droplet generator 606 is shown in 6 and Fig. 7, but more than one such droplet generator 606 can be to device 600 Microfluidic circuit with 700 provides drop.Droplet generator 606 itself may include electrowetting configuration, and therefore can wrap It includes：Substrate with photoresponsive layer may include a-Si：H (for example, as shown in the 6th, 958, No. 132 United States Patent (USP))；Light Cause driving circuit substrate (such as shown in U.S. Patent application of Publication No. 2014/0124370))；Based on photoelectric crystal The substrate (for example, as shown in the 7th, 956, No. 339 United States Patent (USP)) of pipe；Or electric actuation circuitry substrate (for example, such as the 8th, 685, Shown in No. 344 United States Patent (USP)s).Alternatively, droplet generator can have T shapes or Y shape fluid dynamics structure (for example, such as the 7th, No. 708,949, the 7,041,481st (being published as RE41,780 again) number, No. 2008/0014589, the 2008/0003142nd Number, shown in No. 2010/0137163 and No. 2010/0172803 U.S. patents and patent applications disclosure).It is all above-mentioned The full content of american documentation literature is incorporated herein by reference.

As shown, droplet generator 606 may include one or more fluids input 602 and 604 (show two, But can have less or more) and fluid output 208, it may be coupled to microfluidic channel 614.Liquid medium 622,624, life Object small items 630, reagent and/or other biological medium can be loaded by input 602 and 604 in droplet generator 606. Droplet generator 606 can generate liquid medium 622 (its can with but include not necessarily one or more biological speck bodies 630) it, the drop 620 of reagent or other biological medium and is output in channel 614.If channel 614 is matched with electrowetting It sets, then so that drop 620 is moved in channel 614 using electrowetting (or photoelectricity wetting).Alternatively, can exist by other means Mobile drop 620 in channel 614.It is, for example, possible to use fluid flowing, gravity etc. make drop 620 be moved in channel 614.

As described above, microfluidic channel 614 and selection chamber 616/716 can be filled with first fluid medium 624, and Microfluidic channel 612 and the chamber 616 directly connected can be filled with second fluid medium 622.Second fluid medium 622 (hereinafter referred to as " aqueous medium ") can be aqueous medium, such as the sample for the biological speck body 630 such as maintaining, cultivating is situated between Matter.First fluid medium 624 (hereinafter referred to as " unmixability medium ") can be the unmixing medium of aqueous medium 622.Aqueous Jie Matter 622 and the example of unmixability medium 624 include any example discussed above for various media.

Droplet generator 606 can be used for loading biological speck body and/or promote microfluidic device on biochemistry and/ Or the operation of molecular biology workflow.Fig. 6 and Fig. 7 show non-limiting example.By using droplet generator, the device There can be electrowetting configuration in entire fluid circuit.

Fig. 6 and Fig. 7 is shown in which that droplet generator 606 generates the drop 620 for including reagent (or other biological material) Example.Drop 620 containing reagent may move through microfluidic channel 14 and enter the chamber for including unmixability medium 624 One of room 616/716.It is one or more before or after the drop 620 containing reagent is moved into one of chamber 616/716 One or more speck bodies 630 in drop 632 can be moved in identical chamber 616/716.It is then possible to will contain The drop 620 of reagent merges with the drop 632 comprising speck body 630 so that the content of the reagent and drop 632 of drop 620 The biochemical reaction of hybrid concurrency.As shown in Figure 6 and Figure 7, the drop 632 of one or more bodies containing speck can be occurred by drop Device 606 is supplied (not shown) or can be obtained from holding fence 616.Speck body 630 can be biological speck body, such as carefully Born of the same parents are optionally cultured (for example, in chamber 616) before being moved to processing chamber housing 616/716.Alternatively, speck Body 630 can be bead, such as (such as can have been used to cultivate in specimen material 622 in conjunction with interested molecule in sample The cell secreta being present in after one or more of biological cells in specimen material 622) affine bead.In other replacements In scheme, one or more drops 632 can not contain speck body, and only contain aqueous medium, such as specimen material 622, Such as it contains cell secreta after specimen material 622 has been used to cultivate one or more of biological cells.

Fig. 8 show can include such as the microfluidic device of the microfluidic circuit of any of device 600 and 700 in The example of the process 800 of execution.

In the step 802 of process 800, it can be trained in the holding fence full of sample media (such as cell culture medium) Health object speck body.For example, the speck body 630 of Fig. 6 or Fig. 7 can be biology and can be cultivated in its chamber 616.It is logical It can often be cultivated as described above.For example, culture may include using 622 perfusion channel 612 of culture medium.Step 802 can refer to It is executed in the fixed period.

In step 804, the biological speck body of culture can be moved to from sample media filled chamber 616 and be filled with sample The chamber 616/671 of the unmixing medium of product medium, wherein biological speck body is cultivated in sample media filled chamber 616.Example Such as, as described above, the speck body 630 of culture can be in the drop 620 or 632 of sample media 622 from a holding fence 616 It is moved to one to keep in fence 616/716, as shown in Figure 6 and Figure 7.

In step 806, one can be carried out to the biological speck body of culture in the holding fence of unmixability media filler Kind or more processing or process.For example, one or more drops 620 containing one or more of reagents can be sent out by drop Raw device 606 generates, and moves into unmixability media filler chamber 612/716, and with the drop of the biological speck body 630 containing culture 632 merge, and as shown in Figure 6 and Figure 7 and are discussed above.For example, first can contain lytic agent containing reagent droplet 620.Contain There is the drop 632 of the biological speck body 630 of culture that will cause to cultivate with the merging containing reagent droplet 620 of first containing lytic agent Biological speck body 630 dissolving.In other words, it will be formed and combine drop (not shown), it includes the biology from culture is micro- The cell lysate of object 630.Then can by the drop 620 containing other (such as second, third, 4th etc.) reagents with contain The new droplet coalescence of cell lysate, to be further processed cell lysate as needed.

Additionally or as another example, one or more capture speck bodies containing one or more labels are (not Show) drop can be generated by droplet generator 606 and be moved to the fence 616 or 716 of unmixability media filler In, and the droplet coalescence with the sample medium 622 of the biological speck body 630 containing culture in a similar way, the capture are micro- Interested secretion that object generates the biological speck body 630 of culture or one or more substances (such as nucleic acid, such as DNA or RNA, protein, metabolin or other biological molecule) there is affinity.It has been split in the biological speck body 630 of culture In the case of solution, including the drop 620 of capture speck body can be comprising one or more affine beads (for example, to such as The nucleic acid such as DNA, RNA, Microrna have affinity), with keep fence 616 or 716 in the liquid containing cell lysate Drop can be incorporated into the target molecule being present in lysate when merging.

In step 808, processed biological speck body can be optionally processed.For example, if in step 806, will capture Object (not shown) moves into the chamber 616/716 of unmixability media filler with the biological speck body 630 of culture together, then can be with In step 808 chamber 616/716, the capture speck body of reaction instruction and label are monitored for reaction (for example, fluorescence signal) In conjunction with interested material amount.Alternatively, it is micro- to remove (for example, in drop 622) such capture from chamber 616/716 Object (not shown), and output it from microfluidic device (not showing in figure 6 and figure 7) for subsequent analysis.As Processed biological speck body 630 can be removed (for example, in drop 632) simultaneously by another example from chamber 616/716 And it is used for subsequent analysis from the output of microfluidic device (not shown).

Fig. 9 outlines the substrate for being used to form the microfluidic device including electrowetting configuration and dielectrophoresis (DEP) configuration Method.For example, the method described in Fig. 9 can be used to form the integral type lining bottom of type shown in the microfluidic device of Fig. 5.Figure 10-18 depicts the sectional view of the intermediate structure formed after each step in the method for performing Fig. 9.With including light The substrate of the DEP configurations of electric transistor array is the starting point in Figure 10-18.Certainly, as it will appreciated by a person of ordinary skill, rising Beginning substrate is not limited to the substrate with the DEP configurations of photo-transistor arrays, but can be applied to other kinds of substrate, all It such as include the substrate of the electrod-array of amorphous silicon layer or electric actuation.In addition, the step in the method for Fig. 9 can be used alone and/ Or used with other combinations, to generate the other kinds of microfluidic device with conductive substrates, including retouch herein Other microfluidic devices stated.

Step 902 in the method for Fig. 9 includes the initial substrate prepared for further processing.Such as vertical section in Figure 10 Shown in face, initial substrate 1000 includes the high doped layer of conductive silicon 1010, has been formed with photo-transistor arrays on it 1020.The step of preparing substrate 1000 may include thermal anneal process.The process of step 902 can be with the surface of preparing substrate 1000 To ensure the appropriate combination for the material being subsequently deposited on substrate 1000.

Step 904 in the method for Fig. 9 includes the depositing selective anticorrosive additive material on the top surface of initial substrate.Such as figure Shown in vertical cross-section in 11, on the top surface of substrate 1000 deposit one layer of condition erosion resistant 1130 so that its cover at The surface of the phototransistor 1020 of array.In some embodiments, condition erosion resistant 1130 can be nitride.

Step 906 in the method for Fig. 9 includes that the first pattern is applied to the item being deposited on substrate during step 904 On part erosion resistant.As shown in figure 12, which allows in selection area (for example, the phototransistor battle array in 1000 left side of substrate The surface of row) in from substrate 1000 remove condition erosion resistant 1130.It is well known that it will figure such as in industry Case is applied to the condition erosion resistant 1130 being deposited on during step 904 on substrate 1000 can be by photoetching process come real It is existing.This photoetching process includes such as electron beam, X-ray, UV and deep frequency UV.Typically, the pattern is limited using polymer.

As described in the step 908 of the method for Fig. 9, then by depositing photoresponsive layer on pattern then selectively by portion Response layer exposure (for example, there is the light of suitable wavelength and intensity for the material of photoresponsive layer) is divided to process in step The pattern (for example, polymer) deposited in 906.

Step 910 in the method for Fig. 9 include by photoresponsive layer (and positioned at photoresponsive layer etchable beneath portions appoint What condition erosion resistant) it is etched down to the first precalculated position.As shown in figure 12, the first precalculated position can be such as substrate Surface (for example, surface of phototransistor 1020).

Optional subsequent step set (not shown) in the method for Fig. 9 is once substrate is relative to condition resist layer Patterning, then deposit, pattern and etch layer of conductive material.As shown in figure 13, conductive material 1330 can be deposited on substrate Item surface (for example, surface of the phototransistor 1020 in 1000 left side of substrate) and be not removed during step 908 and 910 On the part of part resist layer 1130.Conductive material 1330 can be such as conductive silicon, such as non-crystalline silicon or highly doped silicon.Then, As shown in figure 14, the patterning of conductive material 1330 and etching can cause substrate 1000 first part have be deposited directly to Layer of conductive material 1330 (for example, on surface of the phototransistor 1020 in 1000 left side of substrate) thereon, and substrate 1000 second part has the one layer of condition erosion resistant 1130 of Direct precipitation on it (for example, on 1000 right side of substrate On the surface of phototransistor 1020).

Step 912 in the method for Fig. 9 is included in substrate and (or has been deposited on any material not being etched on substrate and Material) on deposit an at least dielectric layer.Such as (for example, in conjunction with equipment of Figure 1B) discussed elsewhere herein, dielectric layer stack Each layer (such as first layer dielectric material, second layer dielectric material, third layer dielectric material etc.) can be sequentially deposited On substrate.For example, as shown in figure 15, the dielectric stack 1530 being made of two layers of dielectric materials can be deposited on substrate 1000 On.In order to consistent with the other parts of this paper, the first layer of dielectric stack 1530, which needs not be, to be deposited on substrate 1000 First layer.On the contrary, since term first and second can be arbitrarily using or about surface and be moved into substrate The sequence of dielectric materials layer uses.Therefore, in the context of Figure 15, the first layer dielectric material being deposited on substrate 1000 can To be " second layer " dielectric material, and the second layer dielectric material being deposited on substrate 1000 can be " first layer " dielectric material Material.

Step 914 in the method for Fig. 9 includes applying the second pattern on the top of at least one dielectric layer and should At least one dielectric layer etch is to the second precalculated position.In some embodiments, the second precalculated position can be that this layer of condition is anti- The surface of corrosion material 1130.It therefore, as shown in figure 16, can be by each layer of dielectric stack 1530 from the selected part of substrate 1000 The surface until condition erosion resistant 1130 is etched away downwards.As discussed above, condition erosion resistant 1130 can be nitridation Object.Therefore, the etching material used in step 914 can be adapted for etching away dielectric material rather than nitride.

In various embodiments, optional step can further be executed.For example, third pattern can be deposited, and can To execute the stripping of condition resist layer (it can optionally lead to the etching that 10 microns are up to silicon substrate).As shown in figure 17, Condition resist layer 1130 is etched away from the right side of substrate 1000, causes the surface of the phototransistor 1020 on right side sudden and violent again Dew.In addition, as shown in figure 18, can take steps to execute substrate 1000 bottom oxide strips and back metal with One layer of conductive metal 1830 (such as silver or gold) is added to substrate.The substrate obtained shown in Figure 18, which can have, to be configured to It generates the first part (for example, on right side) of DEP power and is configured as generating the second part of electrowetting power (for example, on a left side Side).Joint between the first and second parts, at least with DEP and electrowetting power is generated, substrate can be that electricity is inactive (electrically inactive).The thickness of not active region, and can by the precision depending on mask and etching step To be the thickness (for example, less than 1.5mm, being less than 1.0mm, less than 0.5mm or less) for being, for example, less than 2mm.

Although specific embodiments of the present invention and application have been described in the present specification, these embodiments and answer With being only exemplary, and many variations are possible.For example, can be about the specimen material containing cell secreta The method that (for example, after specimen material 682 has been used to cultivate one or more of biological cells) executes Fig. 8.In this way Embodiment in, step 802 will remain unchanged, but step 804 will be related to that aqueous medium will likely be contained only without speck body The drop 632 of (such as specimen material 622 comprising cell secreta) is moved to the chamber 616/716 of the medium containing unmixability In, and step 806 and 808 will be executed for the drop 632 of this property of water-bearing medium.In addition, the electrowetting being discussed herein is matched It can be any kind of electronics wetting configuration known in the art to set, and example is disclosed in the 6th, 958, No. 132 United States Patent (USP) The U.S. Patent application of (being configured for OEW) and Publication No. US2016/0158748 (for single side OEW configurations).Electrowetting is matched Other examples set include electrowetting (EWOD) equipment on dielectric, can be electronically controlled, one example is It is disclosed in No. 8,685,344 United States Patent (USP)s.Similarly, the dielectrophoresis configuration being discussed herein can be as known in the art any The dielectrophoresis of type configures, and example is disclosed in RE 44, No. 711 (Wu et al.), the 7th, 956, No. 339 (Ohta et al.), the No. 6,294,063 (Becker et al.), No. 6,942,776 (Medoro) and No. 9,403,172 (Wu et al.) United States Patent (USP). The full content of all above-mentioned american documentation literatures is incorporated herein by reference.

Example

System and microfluidic device：For operate it microfluidic device and instrument by Berkeley Lights corporations It makes.The system includes at least flow controller, temperature controller, fluid media (medium) adjusting and pump group part, is used for photoactivation DEP or EW Light source, erecting bed and the camera of configuration.Microfluidic device includes that there is the EW on surface as described below to configure.

Example 1.Preparing has the electrowetting microfluidic equipment of modified inner surface.Using 100W power, 240 millitorr pressure and 440sccm oxygen flows processing microfluidic device (Berkeley in oxygen plasma cleaner (Nordson Asymtek) Lights companies) 1 minute, which has base portion, lid and by base portion and covers separated photo-patterning silicones Microfluidic circuit material, the base portion include the electrode active of the semiconductor layer with photosensitive silicon and the dielectric layer that upper surface is aluminium oxide Change substrate, covering has the glass supports with ITO electrode.Exist in foil boat in the bottom of vacuum reactor and is used as water It is in the case of the bitter salt (0.5g, Acros) of reaction source, plasma treated microfluidic device is anti-in vacuum Answer in device, with trimethoxy (3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,11,11,12,12,13,13,14,14, 15,15,16,16,16- bis- ten nine fluorine cetyls) silane (0.3g, such as on October 19th, 2016 U.S. Provisional Application submitted Synthesis details described in 62/410238) it is handled (in the independent foil boat of the bottom of vacuum reactor).Then, make Cavity pump to 750 millitorrs and is sealed with vacuum pump.Vacuum reactor is placed in be heated in 180 DEG C of stove up to 24-48 it is small When.After being cooled to room temperature and argon gas is introduced vacuum chamber, there is dimethoxy on all inner surfaces from removal in reactor Base (3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,11,11,12,12,13,13,14,1 4,15,15,16,16,16- 29 fluorine cetyls) siloxy moieties outer hydrophobic layer microfluidic device.After removal, silicone oil (5 is being used using preceding Centistoke viscosity, Gelest Cat.#DMS-T05) filling microfluidic device.Figure 20 A-20C are in the silicone oil phase of unmixability The chronophotograph image of mobile water droplet around hydrophobic layer (that is, drop actuation surfaces).It is configured using the electrowetting of optical actuation With the drop actuation surfaces of microfluidic device, drop shows excellent locomotivity.

The reproduction of embodiment

1. a kind of microfluidic device with electrowetting configuration, the microfluidic device include：

Substrate has dielectric layer, drop actuation surfaces and the first electrode for being configured to connect to AC voltage sources；And

Second electrode is configured to connect to the AC voltage sources；

The wherein described dielectric layer is electrically coupled to the first electrode, also,

The wherein described drop actuation surfaces include being covalently bound to the hydrophobic layer of the dielectric layer.

2. according to microfluidic device described in embodiment 1, wherein there is the equipment unilateral electrowetting to configure.

3. according to the microfluidic device described in embodiment 2, wherein the second electrode be made of the substrate it is netted Electrode.

4. according to microfluidic device described in embodiment 1, wherein the equipment has photoelectricity wetting (OEW) configuration.

5. according to microfluidic device described in embodiment 1, wherein there is the equipment electrowetting (EWOD) on dielectric to match It sets.

6. the microfluidic device according to any one of embodiment 1 to 5, wherein the hydrophobic layer is to include that surface is modified Ligand and the surface modified ligand is linked to the surface the link group single layer, wherein the drop actuation surfaces have There is the structure of Formula II：

WhereinIt is the surface of dielectric layer；V is-P (O) (OY) W- or-Si (OZ)₂W-；W is-O- ,-S- or-NH- and connects To surface；Z is the key to the adjacent silicon atoms for being attached to surface or the key to surface；Y is to the adjacent phosphorus for being attached to surface The key of atom or key to surface；R is hydrogen or fluorine；M is hydrogen or fluorine；H is independently 2 or 3 integer；J is 1；K is 0 or 1；m For 0 or integer of 1 to 20；N is 0 or integer of 1 to 20；The integer that the summation of (n+ [(h+j) k]+m) is 11 to 25；When k is When 1, then it is hydrogen that m, which is at least 2 and M,；When k is 0 and R is fluorine, then it is hydrogen that m, which is at least 2 and M,.

7. the microfluidic device according to any one of embodiment 1 to 6, wherein the electrowetting configuration of the equipment includes The first part of the equipment, and the wherein described equipment further includes the second part that there is dielectrophoresis (DEP) to configure.

8. a kind of microfluidic device, including：Substrate has at least one electrode for being configured to connect to voltage source；Lid, With at least one electrode for being configured to connect to the voltage source；And at least one spacer element,

The wherein described substrate and the lid are substantially parallel to each other and are bonded together by the spacer element to limit It is configured as keeping the capsule of liquid, wherein the substrate has the drop actuation surfaces for partly limiting the capsule, it is described Drop actuation surfaces have inner-dielectric-ayer and outer hydrophobic layer,

The wherein described outer hydrophobic layer includes the self-association molecule on the surface for being covalently bound to the inner-dielectric-ayer, at it The upper hydrophobic monolayer for forming dense packing, and

Wherein when at least one electrode of at least one electrode of the substrate and the lid is connected to the voltage source When opposing terminal, the substrate being capable of pair aqueous drop contacted with the drop actuation surfaces of the substrate application electrowetting Power.

9. according to the microfluidic device described in embodiment 8, wherein the self-association molecule of the hydrophobic monolayer includes each table Face modified ligand and the link group that the surface modified ligand is linked to the inner-dielectric-ayer surface, wherein the drop causes Dynamic surface has the structure of Formula II：

10. according to the microfluidic device described in embodiment 9, wherein V is-Si (OZ)₂W-。

11. according to the microfluidic device described in embodiment 9, wherein V is-P (O) (OY) W-.

12. the microfluidic device according to any one of embodiment 9 to 11, wherein n are integer of 1 to 20, and its Middle R is hydrogen.

13. according to the microfluidic device described in embodiment 12, wherein m is integer of 1 to 20, and wherein M is hydrogen.

14. according to the microfluidic device described in embodiment 13, wherein m is 2.

15. the microfluidic device according to any one of embodiment 9 to 11, wherein n are integer of 1 to 20, and its Middle R is fluorine.

16. according to the microfluidic device described in embodiment 15, wherein m is integer of 1 to 20, and wherein M is hydrogen.

17. according to the microfluidic device described in embodiment 16, wherein m is 2.

18. the microfluidic device according to any one of embodiment 9 to 17, wherein k are 1.

19. the microfluidic device according to any one of embodiment 9 to 17, wherein k are 0.

20. the microfluidic device according to any one of embodiment 9 to 19, wherein the summation of (n+ [(h+j) k]+m) For 13 to 19 integer.

21. the microfluidic device according to any one of embodiment 8 to 20, wherein the drop actuating of the substrate The outer hydrophobic layer on surface has the thickness less than 5 nanometers.

22. the microfluidic device according to any one of embodiment 8 to 21, wherein the drop actuating of the substrate The outer hydrophobic layer on surface is patterned such that compared with the rest part of the outer hydrophobic layer, selection area is relatively close Water.

23. the microfluidic device according to any one of embodiment 8 to 22, wherein the drop actuating of the substrate The inner-dielectric-ayer on surface includes oxidiferous first layer dielectric material.

24. the microfluidic device according to any one of embodiment 8 to 23, wherein the oxide is metal oxidation Object.

25. according to the microfluidic device described in embodiment 24, wherein the metal oxide is aluminium oxide.

26. the microfluidic device according to any one of embodiment 23 to 25, wherein the first layer dielectric material is logical Atomic layer deposition is crossed to be formed.

27. the microfluidic device according to any one of embodiment 23 to 26, wherein the drop of the substrate causes The inner-dielectric-ayer on dynamic surface further includes second layer dielectric material, and the wherein described outer hydrophobic layer is covalently bound to described the One layer of dielectric material.

28. according to the microfluidic device described in embodiment 27, wherein second layer dielectric material includes oxide or nitride.

29. according to the microfluidic device described in embodiment 28, wherein second layer dielectric material is selected from silica and nitridation The group that silicon is constituted.

30. the microfluidic device according to any one of embodiment 27 to 29, wherein the second layer dielectric material is logical Plasma enhanced chemical vapor deposition is crossed to be formed.

31. the microfluidic device according to any one of embodiment 23 to 30, wherein the first layer dielectric material packet The first sublayer dielectric material and the second sublayer dielectric material are included, wherein first sublayer is covalently bound to the hydrophobic layer.

32. according to the microfluidic device described in embodiment 31, wherein the first sublayer dielectric material includes silica.

33. according to the microfluidic device described in embodiment 31, wherein the first sublayer dielectric material passes through ALD deposition.

34. the microfluidic device according to any one of embodiment 31 to 33, wherein the first layer dielectric material has There is about 10nm to the thickness of about 20nm.

35. according to the microfluidic device described in embodiment 34, wherein the first sublayer dielectric material has about 2nm to about The thickness of 10nm.

36. the microfluidic device according to any one of embodiment 8 to 35, wherein the drop actuating of the substrate The inner-dielectric-ayer on surface has at least about 40 nanometers of thickness.

37. according to the microfluidic device described in embodiment 36, wherein the drop actuation surfaces of the substrate is described Inner-dielectric-ayer has about 40 nanometers to about 120 nanometers of thickness.

38. the microfluidic device according to any one of embodiment 8 to 37, wherein the substrate further includes photoresponse Layer, the photoresponsive layer have the first side for contacting the inner-dielectric-ayer and contact the second side of at least one electrode.

39. according to the microfluidic device described in embodiment 38, wherein the photoresponsive layer includes amorphous silicon hydride (a-Si： H)。

40. the microfluidic device according to embodiment 38 or 39, wherein the photoresponsive layer is at least 900 nanometers Thickness.

41. according to the microfluidic device described in embodiment 40, wherein the photoresponsive layer is with about 900 to 1100 nanometers Thickness.

42. according to the microfluidic device described in embodiment 38, wherein the photoresponsive layer includes multiple conductors, each conductor At least one electrode of the substrate is controllably connected to via phototransistor switch.

43. the microfluidic device according to any one of embodiment 8 to 42, wherein the substrate includes the company of being configured as It is connected to the single electrode of AC voltage sources, the single electrode includes tin indium oxide (ITO) layer.

44. the microfluidic device according to any one of embodiment 8 to 42, wherein the substrate includes the company of being configured as It is connected to the single electrode of AC voltage sources, the single electrode includes conductive silicon layer.

45. the microfluidic device according to any one of embodiment 8 to 37, wherein the substrate includes multiple electrodes, Each electrode is configured to connect to one or more AC voltage sources.

46. according to the microfluidic device described in embodiment 45, wherein each electrode in the multiple electrode is via crystal Pipe switch may be connected to one in one or more AC voltage sources.

47. the microfluidic device according to any one of embodiment 8 to 46, wherein the lid, which has, partly limits institute The inner surface of capsule is stated, the inner surface of the lid has internal layer and outer hydrophobic layer, wherein the lid is described outer thin Water layer includes the self-association molecule on the surface for the internal layer for being covalently bound to the lid, to be formed on dense packing Hydrophobic monolayer.

48. according to the microfluidic device described in embodiment 47, wherein the self-association molecule of the hydrophobic monolayer of the lid is each The link group on the surface of the internal layer of the lid is linked to including surface modified ligand and by the surface modified ligand, Described in the inner surface that covers there is the structure of Formula II：

WhereinIt is the surface of dielectric layer；V is-P (O) (OY) W- or-Si (OZ)₂W-；W is-O- ,-S- or-NH- and connects To surface；Z is the key to the adjacent silicon atoms for being attached to surface or the key to surface；Y is to the adjacent phosphorus for being attached to surface The key of atom or key to surface；R is hydrogen or fluorine；M is hydrogen or fluorine；H is independently 2 or 3 integer；J is 1；K is 0 or 1；m For 0 or integer of 1 to 20；N is 0 or integer of 1 to 20；The integer that the summation of (n+ [(h+j) k]+m) is 11 to 25；When k is When 1, then it is hydrogen that m, which is at least 2 and M,；And when k is 0 and R is fluorine, then it is hydrogen that m, which is at least 2 and M,.

49. according to the microfluidic device described in embodiment 48, wherein the self-association molecule of the hydrophobic monolayer of the lid and institute The self-association molecule for stating the hydrophobic monolayer of the drop actuation surfaces of substrate is identical.

50. the microfluidic device according to any one of embodiment 47 to 49, wherein the inner surface of the lid is outer Hydrophobic layer has the thickness less than 5 nanometers.

51. the microfluidic device according to any one of embodiment 47 to 50, wherein the internal layer of the lid is interior dielectric Layer.

52. according to the microfluidic device described in embodiment 51, wherein the lid further includes photoresponsive layer.

53. according to the microfluidic device described in embodiment 51, wherein the lid includes multiple electrodes, each electrode is configured To be connected to one or more AC voltage sources.

54. according to the microfluidic device described in embodiment 8, wherein at least one spacer element includes that silicon substrate is organic poly- Close object.

55. according to the microfluidic device described in embodiment 54, wherein the silicon substrate organic polymer is selected from poly dimethyl silicon The group that oxygen alkane (PDMS) and light patternable polysiloxanes (PPS) are constituted.

56. the microfluidic device according to any one of embodiment 8 to 53, wherein at least one spacer element packet Include SU-8.

57. the microfluidic device according to any one of embodiment 8 to 56, wherein at least one spacer element tool There is at least 30 microns of thickness.

58. the microfluidic device according to any one of embodiment 8 to 57, wherein at least one spacer element exists One or more microchannels are limited in the capsule.

59. according to the microfluidic device described in embodiment 58, wherein at least one spacer element is in the capsule Multiple chambers are further limited, wherein each chamber is from least one microchannel openings.

60. a method of manufacture microfluidic device, the method includes：Spacer element is attached to have and is configured as It is connected to the inner surface of the lid of at least one electrode of voltage source；

The spacer element and lid are attached to the substrate at least one electrode for being configured for connection to voltage source Dielectric surface, thus the spacer element become to be arranged between the inner surface and the dielectric surface of the substrate of the lid, The lid and the substrate are oriented substantially parallel to one another, and the substrate, spacer element and common limit of lid are configured to Keep the capsule of liquid；The hydrophobic list of dense packing is formed at least part of the inner surface of the lid by being vapor-deposited Layer, wherein the hydrophobic monolayer includes the self-association molecule for the inner surface for being covalently bound to the lid；And pass through vapor deposition The hydrophobic monolayer of dense packing is formed at least part of the dielectric surface of the substrate, wherein the hydrophobic monolayer includes It is covalently bound to the self-association molecule of the dielectric surface of the substrate.

61. according to the method described in embodiment 60, wherein the self-association molecule and the substrate of the hydrophobic monolayer of the lid The self-association molecule of hydrophobic monolayer include each surface modified ligand and be linked to the surface modified ligand respectively described The link group of the inner surface of lid and the dielectric surface of substrate, wherein the gained surface of the lid and the substrate has Formula II Structure：

62. according to the method described in embodiment 61, wherein V is-Si (OZ)₂W-。

63. according to the method described in embodiment 61, wherein V is-P (O) (OY) W-.

64. the method according to any one of embodiment 61 to 63, wherein n are integer of 1 to 20, and wherein R is Hydrogen.

65. according to the method described in embodiment 64, wherein m is integer of 1 to 20, and wherein M is hydrogen.

66. according to the method described in embodiment 65, wherein m is 2.

67. the method according to any one of embodiment 61 to 63, wherein n are integer of 1 to 20, and wherein R is Fluorine.

68. according to the method described in embodiment 67, wherein m is integer of 1 to 20, and wherein M is hydrogen.

69. according to the method described in embodiment 68, wherein m is 2.

70. the method according to any one of embodiment 61 to 69, wherein k are 1.

71. the method according to any one of embodiment 61 to 69, wherein k are 0.

72. the microfluidic device according to any one of embodiment 61 to 71, wherein (n+ [(h+j) k]+m's) is total With the integer for 13 to 19.

73. a kind of microfluidic device, including：Conductive silicon substrate has dielectric stack and is configured to connect to voltage source At least one electrode；Lid has at least one electrode for being configured to connect to voltage source；And at least one interval member Part,

The wherein described conductive silicon substrate and the lid are substantially parallel to each other and are bonded on one by the spacer element It rises to limit the capsule for being configured as keeping liquid,

The wherein described conductive silicon substrate has the inner surface for partly limiting the capsule, described inwardly facing surface including institute The outmost surface of dielectric stack is stated, and wherein when at least one electrode of at least one electrode of the substrate and the lid When being connected to the opposing terminal of AC voltage sources, the substrate being capable of pair aqueous drop contacted with the inner surface of substrate application electricity Wetting power.

74. according to the microfluidic device described in embodiment 73, wherein the conductive silicon substrate includes non-crystalline silicon.

75. according to the microfluidic device described in embodiment 73, wherein the conductive silicon substrate includes photo-transistor arrays.

76. according to the microfluidic device described in embodiment 73, wherein the conductive silicon substrate includes electrod-array.

77. the microfluidic device according to any one of embodiment 73 to 76, wherein the conductive silicon substrate is inside Surface further includes outer hydrophobic layer, and the outer hydrophobic layer includes the self-association molecule for being covalently bound to the interior dielectric stack.

78. the microfluidic device according to any one of embodiment 73 to 77, wherein the interior dielectric stack includes First layer dielectric material and second layer dielectric material.

79. according to the microfluidic device described in embodiment 78, wherein first layer dielectric material has first surface and opposite Surface, wherein the first surface of the first layer abuts the second layer, and the apparent surface of the wherein described first layer is formed The outmost surface of the dielectric stack.

80. the microfluidic device according to embodiment 78 or 79, wherein the first layer dielectric material includes metal oxygen Compound.

81. according to the microfluidic device described in embodiment 80, wherein the first layer dielectric material includes aluminium oxide or oxygen Change hafnium.

82. the microfluidic device according to any one of embodiment 78 to 81, wherein the second layer dielectric material packet Include oxide or nitride.

83. according to the microfluidic device described in embodiment 82, wherein the second layer dielectric material includes silica or nitrogen SiClx.

84. the microfluidic device according to any one of embodiment 78 to 83, wherein the second layer passes through plasma Body enhances chemical vapor deposition (PECVD) technology to deposit.

85. the microfluidic device according to any one of embodiment 78 to 84, wherein the first layer passes through atomic layer (ALD) technology of deposition deposition.

86. the microfluidic device according to any one of embodiment 78 to 85, wherein the interior dielectric stack includes Third layer with first surface and apparent surface, wherein the first surface of the third layer abuts the opposite table of the first layer Face, and the apparent surface of the wherein described third layer forms the outmost surface of the dielectric stack.

87. according to the microfluidic device described in embodiment 86, wherein the third layer includes silica.

88. the microfluidic device according to embodiment 86 or 87, wherein the third layer passes through atomic layer deposition (ALD) Technology deposits.

89. the microfluidic device according to any one of embodiment 78 to 85, wherein the first layer dielectric material has There is about 10nm to the thickness of about 50nm.

90. the microfluidic device according to any one of embodiment 86 to 88, wherein the first layer dielectric material has There is about 5nm to the thickness of about 20nm, and the third layer dielectric material has the thickness of about 2nm to about 10nm.

91. the microfluidic device according to any one of embodiment 78 to 90, wherein the second layer dielectric material has There is about 30nm to the thickness of about 100nm.

92. the microfluidic device according to any one of embodiment 73 to 91, wherein the drop of the substrate activates table The dielectric stack in face has at least about 40 nanometers of thickness.

93. according to the microfluidic device described in embodiment 92, wherein the dielectric heap of the drop actuation surfaces of the substrate The folded thickness with about 40 nanometers to about 120 nanometers.

94. the microfluidic device according to any one of embodiment 73 to 93, wherein the dielectric layer has the Europe about 50k The impedance of nurse to about 150k ohm.

95. the microfluidic device according to any one of embodiment 73 to 94, wherein described device include：

Dielectrophoresis module, for applying voltage in response to first under first frequency to execute the first microfluidic procedures；With And

Electrowetting module, for receiving the output from the dielectrophoresis module, and in response to the under second frequency Two apply voltages to execute the second microfluidic procedures,

The wherein described Electrowetting module includes the dielectric stack of the conductive silicon substrate.

Further include the bridge between the first module and the second module 96. according to the microfluidic device described in embodiment 95.

97. according to the microfluidic device described in embodiment 96, wherein the bridge do not execute first microfluidic procedures or Second microfluidic procedures.

98. the microfluidic device according to embodiment 96 or 97, wherein the bridge is charge neutrality.

99. the microfluidic device according to any one of embodiment 96 to 98, wherein the bridge includes pipeline.

100. the microfluidic device according to any one of embodiment 96 to 98, wherein the bridge includes polymer.

101. the microfluidic device according to any one of embodiment 95 to 100, wherein the output is biomaterial.

102. the microfluidic device according to any one of embodiment 95 to 101, wherein the first frequency exists In the range of 100kHz to 10mHz.

103. the microfluidic device according to any one of embodiment 95 to 102, wherein the second frequency is in 1kHz To 300kHz.

104. the microfluidic device according to any one of embodiment 95 to 103, wherein the first voltage is 1 to 10 In the range of volt.

105. the microfluidic device according to any one of embodiment 95 to 104, wherein the second voltage 10 to In the range of 100 volts.

106. the microfluidic device according to any one of embodiment 95 to 105, wherein the conductive silicon substrate is whole Body.

107. the microfluidic device according to any one of embodiment 95 to 106, wherein the conductive silicon substrate is double Body.

108. according to the microfluidic device described in embodiment 106, wherein the conductive silicon substrate includes non-crystalline silicon.

109. according to the microfluidic device described in embodiment 107, wherein the conductive silicon substrate includes non-crystalline silicon.

110. according to the microfluidic device described in embodiment 106, wherein the conductive silicon substrate includes phototransistor battle array Row.

111. according to the microfluidic device described in embodiment 107, wherein the conductive silicon substrate includes phototransistor battle array Row.

112. according to the microfluidic device described in embodiment 106, wherein the conductive silicon substrate includes electrod-array.

113. according to the microfluidic device described in embodiment 107, wherein the conductive silicon substrate includes electrod-array.

114. it is a kind of be used for transmission speck body that is compatible with aqueous medium and/or dissolving in aqueous medium, biological product and/ Or the system of reagent, the system comprises：

Microfluidic device, with the capsule for including base portion and microfluidic circuit structure, wherein the base portion includes covalently tying Close at least part of hydrophobic monolayer of the upper surface of the base portion；

The first fluid medium unmixing with aqueous medium；And

At least one aqueous drop.

115. according to the system described in embodiment 114, wherein the hydrophobic monolayer has surface modified ligand and by surface Modified ligand links to the link group on surface, and wherein hydrophobic surface has the structure of Formula II：

WhereinIt is surface；V is-P (O) (OY) W- or-Si (OZ) 2W-；W is-O- ,-S- or-NH- and is connected to table Face；Z is the key to the adjacent silicon atoms for being attached to surface or the key to surface；Y is to the adjacent phosphorus atoms for being attached to surface Key or key to surface；R is hydrogen or fluorine；M is hydrogen or fluorine；H is independently 2 or 3 integer；J is 1；K is 0 or 1；M is 0 Or integer of 1 to 20；N is 0 or integer of 1 to 20；The integer that the summation of (n+ [(h+j) k]+m) is 11 to 25；When k is 1 When, then it is hydrogen that m, which is at least 2 and M,；And when k is 0 and R is fluorine, then it is hydrogen that m, which is at least 2 and M,.

116. the system according to embodiment 114 or 115, wherein the base portion includes conductive substrates.

117. the system according to any one of embodiment 114 to 116, wherein the microfluidic device is according to implementation Microfluidic device described in any one of example 1 to 59.

118. according to the system described in embodiment 117, wherein the microfluidic device includes the EW configurations of optical actuation.

119. the system according to embodiment 117 or 118, wherein the microfluidic device further includes DEP configurations.

120. the system according to any one of embodiment 114 to 119, wherein the first fluid medium includes having At least one organic compound of backbone structure or at least one organo-silicon compound, the backbone structure include selected from carbon, silicon With the atom of oxygen.

121. according to the system described in embodiment 120, wherein the backbone structure packet of at least one organo-silicon compound Include silicon atom and optional oxygen atom.

122. according to the system described in embodiment 120, wherein the backbone structure of at least one organic compound includes Carbon atom and optional oxygen atom.

123. according to the system described in embodiment 122, wherein the backbone structure is branch.

124. the system according to any one of embodiment 120 to 123, wherein the first fluid medium includes one kind Or more acyclic organic or organo-silicon compound.

125. according to the system described in embodiment 124, wherein the first fluid medium is by acyclic organic or siliconated Close object composition.

126. the system according to any one of embodiment 114 to 125, wherein the first fluid medium does not include complete Fluorinated carbon atom.

127. the system according to any one of embodiment 114 to 125, wherein the compound of the first fluid medium The substituent group of carbon atom include fluoro substituents no more than 90%.

128. the system according to any one of embodiment 115 to 125, wherein the surface modified ligand includes at least First part, the first part include perfluocarbon atom in the end towards inside of the hydrophobic monolayer.

129. according to the system described in embodiment 128, wherein whole carbon atoms of the hydrophobic monolayer are fluoridized.

130. the system according to any one of embodiment 114 to 129, wherein the first fluid medium includes being more than A kind of organic or organo-silicon compound.

131. the system according to any one of embodiment 114 to 130, wherein the capsule further includes lid.

132. according to the system described in embodiment 131, wherein the lid is transparent to light.

133. the system according to embodiment 131 or 132, wherein the lid includes glass and/or indium oxide tantalum (ITO)。

134. the system according to any one of embodiment 131 to 133, wherein the lid includes electrode.

135. the system according to any one of embodiment 114 to 134, wherein aqueous drop includes surfactant.

136. according to the system described in embodiment 135, wherein the surfactant includes nonionic surfactant.

137. the system according to embodiment 135 or 136, wherein the surfactant includes the copolymerization of block oxyalkylene Object, aliphatic ester ethoxylation dehydrated sorbitol, ethoxylation fluorine-containing surfactant, lauryl sodium sulfate or 2,4,7, 9- tetramethyl -5- decine -4,7- diol ethoxylates.

138. the system according to any one of embodiment 135 to 137, wherein the surfactant includesFS-30 (DuPontTM, Synquest Laboratories).

139. the system according to any one of embodiment 114 to 139, wherein the drop includes phosphate buffer salt Solution.

140. the system according to any one of embodiment 114 to 139, wherein aqueous solution drop includes at least one Speck body.

141. system according to embodiment 140, wherein the speck body is biological speck body.

142. system according to any one of embodiment 114 to 141, wherein aqueous drop include containing nucleic acid and/ Or the biological product of protein.

143. system according to any one of embodiment 114 to 142, wherein aqueous drop includes reagent.

144. it is a kind of be used for transmission speck body that is compatible with aqueous medium and/or dissolving in aqueous medium, biological product and/ Or the kit of reagent, the kit include：

Microfluidic device, with the capsule for including base portion and microfluidic circuit structure, wherein the base portion includes covalently tying Close at least part of hydrophobic monolayer of the upper surface of the base portion；And it is situated between with the unmixing first fluid of aqueous medium Matter.

145. kit according to embodiment 144, wherein hydrophobic monolayer have surface modified ligand and change surface Property ligand link to the link group on surface, wherein hydrophobic surface has the structure of Formula II：

WhereinIt is surface；V is-P (O) (OY) W- or-Si (OZ) 2W-；W is-O- ,-S- or-NH- and is connected to surface；Z It is the key to the adjacent silicon atoms for being attached to surface or the key to surface；Y is the key to the adjacent phosphorus atoms for being attached to surface Or the key to surface；R is hydrogen or fluorine；M is hydrogen or fluorine；H is independently 2 or 3 integer；J is 1；K is 0 or 1；M be 0 or 1 to 20 integer；N is 0 or integer of 1 to 20；The integer that the summation of (n+ [(h+j) k]+m) is 11 to 25；When k is 1, then m At least 2 and M is hydrogen；When k is 0 and R is fluorine, then it is hydrogen that m, which is at least 2 and M,.

146. kit according to embodiment 144 or 145, wherein the base portion includes conductive substrates.

147. kit according to any one of embodiment 144 to 146, wherein the microfluidic device is according to reality Apply the microfluidic device described in any one of example 1 to 59.

A kind of 148. processes operating the microfluidic device according to any one of embodiment 8 to 59, the process packet It includes：

The capsule or part of it are filled with the first liquid medium；

Apply AC voltage potentials between at least one electrode of substrate and at least one electrode of lid；

First liquid drop is introduced into the capsule, wherein first drop is immiscible in first liquid medium In；And

By applying electrowetting power to first drop, first drop is moved to the expectation position in the capsule It sets.

149. process according to embodiment 148, wherein first liquid medium is oil.

150. process according to embodiment 148, wherein first liquid medium is silicone oil, fluorinated oil or its group It closes.

151. process according to any one of embodiment 148 to 150, applied in AC voltage potentials be at least 20ppV。

152. process according to embodiment 151, wherein the AC voltage potentials applied are between about 25 and 35ppV.

153. process according to any one of embodiment 148 to 152, applied in AC voltage potentials have about 1 to 100kHz frequency.

154. process according to any one of embodiment 148 to 153, wherein the microfluidic device includes drop life It grows up to be a useful person, and first drop is introduced the capsule by the wherein described droplet generator.

155. process according to any one of embodiment 148 to 154, wherein the first drop packet aqueous solution.

156. process according to embodiment 155, wherein the first drop includes at least one speck body.

157. process according to embodiment 156, wherein at least one speck body is biological speck body.

158. process according to embodiment 157, wherein the biology speck body is cell.

159. process according to any one of embodiment 155 to 158, wherein the aqueous solution is cell culture Base.

160. process according to embodiment 156, wherein at least one speck body is that have to interested material There is the capture bead of affinity.

161. process according to embodiment 160, wherein first drop includes 2 to 20 capture beads.

162. process according to embodiment 160, wherein interested material is biological cell secretion.

163. process according to embodiment 160 or 161, wherein interested material be selected from by DNA, genomic DNA, Mitochondrial DNA, RNA, mRNA, miRNA or any combination thereof composition group.

164.164. the process according to embodiment 155 or 156, wherein first drop includes reagent.

165. process according to embodiment 164, wherein the reagent is cell cracking agent.

166. process according to embodiment 165, wherein the reagent includes non-ionic detergent.

167. process according to embodiment 166, wherein the concentration of the non-ionic detergent is less than 0.2%.

168. process according to embodiment 164, wherein the reagent is proteolytic enzyme.

169. process according to embodiment 168, wherein proteolytic enzyme can be deactivated.

170. process according to any one of embodiment 148 to 169 further includes：

Second drop is introduced into the capsule, wherein the liquid of second drop in first liquid medium not It is miscible but miscible with the liquid of first drop；

By the way that electrowetting power is applied to second drop, second drop is moved to and the first drop phase Position in adjacent capsule；And

Merge second drop and combines drop with first drop to form first.

171. process according to embodiment 170, wherein by being applied to second drop and/or first drop Power-up wetting power makes second drop and first droplet coalescence.

172. process according to embodiment 170 or 171, wherein first drop includes biological speck body, and Wherein described second drop includes reagent.

173. process according to embodiment 172 buffers wherein the reagent for including in second drop is selected from cracking The group that liquid, fluorescent marker and luminescence assays reagent are constituted.

174. process according to embodiment 172, wherein the reagent for including in the second drop is lysis buffer, and The wherein described biological cell is cleaved in first drop and second droplet coalescence.

175. process according to any one of embodiment 170 to 174 further includes：

Third drop is introduced into capsule, wherein the liquid of third drop it is unmixing in first liquid medium but with The liquid of the first combination drop is miscible；And by the way that electrowetting power is applied to the third drop, by the third liquid Drop is moved in the capsule and combines the adjacent position of drop with described first；And

Merge the third drop and combines drop with described first to form the second combination drop.

176. process according to embodiment 175, wherein by the third drop and/or the first combination liquid Drop, which applies electrowetting power, makes the third drop combine droplet coalescence with described first.

177. process according to embodiment 175 or 176, wherein the third drop includes reagent.

178. process according to embodiment 177, wherein the third drop includes protease inhibitors.

179. process according to embodiment 177, wherein the third drop includes having parent to interested material Bead is captured with one to 20 of power.

180. process according to embodiment 179, wherein capture bead includes oligonucleotides capturing agent.

181. process according to embodiment 180, wherein the oligonucleotides capturing agent is poly- dT oligonucleotides.

182. process according to any one of embodiment 179 to 181, wherein the interested material be selected from by DNA, genomic DNA, mitochondrial DNA, RNA, mRNA, miRNA or any combination thereof constitute group.

183. process according to any one of embodiment 179 to 182 further includes：

One to 20 capture bead is exported from the microfluidic device.

184. according to the process according to any one of embodiment 175 to 183, further includes：

4th drop is introduced into the capsule, wherein the liquid of the 4th drop in first liquid medium not Liquid miscible but that drop is combined with described second is miscible；

By the way that electrowetting power is applied to the 4th drop, the 4th drop is moved to and combines drop phase with described second Position in adjacent capsule；And

Merge the 4th drop and combines drop with described second to form third combination drop.

185. process according to embodiment 184, wherein by the 4th drop and/or the second combination liquid Drop, which applies electrowetting power, makes the 4th drop combine droplet coalescence with described second.

186. process according to embodiment 184 or 185, wherein the 4th drop includes reagent.

187. process according to embodiment 186, wherein the reagent for including in the 4th drop includes containing buffering Liquid, dNTP and be adapted for carrying out reverse transcription reaction polymerase mixture.

188. process according to embodiment 186, wherein the reagent for including in the 4th drop includes containing buffering Liquid, dNTP and the mixture for being adapted for the polymerase that whole genome amplification reacts.

189. process according to any one of embodiment 148 to 188, wherein first drop, the second drop, Three drops and the 4th drop are respectively with about 5 to 50 nanoliters of volume.

190. process according to embodiment 189, wherein first drop, second drop and the third liquid Drop is respectively with about 5 to 20 nanoliters of volume.

191. process according to embodiment 190, wherein the volume base of second drop and/or the third drop It is equal to the volume of first drop in sheet.

192. process according to embodiment 190 or 191, wherein the 4th drop is with bigger than first drop About 1 to 3 times of volume.

193. process according to embodiment 192, wherein the 4th drop has about 10 to 30 nanoliters of volume.

194. process according to any one of embodiment 148 to 193, wherein the capsule includes at least one micro- logical Road.

195. process according to embodiment 194, wherein first drop to be moved to the expectation in the capsule Position includes that first drop is moved through at least one microchannel.

196. process according to embodiment 194 or 195, wherein the capsule further includes from described at least one micro- logical Multiple chambers of road opening.

197. process according to embodiment 196, wherein first drop to be moved to the expectation in the capsule Position includes that first drop is moved in the multiple chamber chamber.

198. process according to any one of embodiment 194 to 197, wherein second drop is moved to and institute It includes that second drop is moved through at least one microchannel and is optionally moved to state the adjacent position of the first drop It moves into the chamber for accommodating first drop.

199. process according to embodiment 198 combines liquid wherein being moved to the third drop with described first It includes that the third drop is moved through at least one microchannel and is optionally moved to include to drip adjacent position In the chamber of the first combination drop.

200. process according to embodiment 199, wherein the 4th drop is moved adjacent to the second combination drop Position include by the 4th drop move through at least one microchannel and be optionally into comprising it is described second combination In the chamber of drop.

201. according to the process according to any one of embodiment 148 to 200, wherein applying electrowetting power with movement And/or merge effective electro-wetting property that drop includes the region close to drop for changing the substrate surface.

202. process according to embodiment 201, wherein it includes activating the substrate table to change effective electro-wetting property Electrowetting electrode at the region close to drop in face.

203. process according to embodiment 202, wherein the substrate includes photoresponsive layer, and wherein described in activation Electrowetting electrode at the region close to drop of substrate surface includes that light pattern is directed to described in the electrowetting surface On region.

It is equivalent

Foregoing written specification is considered being enough to enable those skilled in the art to implement these embodiments.The description of front Certain embodiments are detailed with embodiment and describe expected optimal mode.It should be understood, however, that no matter in text How the above to be described in detail, which can put into practice and should be according to appended claims and its any etc. in many ways Jljl is explained.

Claims

Second electrode is configured to connect to the AC voltage sources；

Wherein, the dielectric layer is electrically coupled to the first electrode, also,

Wherein, the drop actuation surfaces include being covalently bound to the hydrophobic layer of the dielectric layer.

2. microfluidic device according to claim 1, wherein there is the equipment unilateral electrowetting to configure.

3. microfluidic device according to claim 2, wherein the second electrode is the netted electricity being made of the substrate Pole.

4. microfluidic device according to claim 1, wherein the equipment has photoelectricity wetting OEW configurations.

5. microfluidic device according to claim 1, wherein the equipment has electrowetting EWOD configurations on dielectric.

6. microfluidic device according to claim 1, wherein the hydrophobic layer be include surface modified ligand and will be described Surface modified ligand links to the single layer of the link group on the surface, wherein the drop actuation surfaces have the knot of Formula II Structure：

Wherein,It is the surface of dielectric layer；

V is-P (O) (OY) W- or-Si (OZ)₂W-；

W is-O- ,-S- or-NH- and is connected to the surface；

Z is the key to the adjacent silicon atoms for being attached to the surface or the key to the surface；

Y is the key to the adjacent phosphorus atoms for being attached to the surface or the key to the surface；

R is hydrogen or fluorine；

M is hydrogen or fluorine；

H is independently 2 or 3 integer；

J is 1；

K is 0 or 1；

M is 0 or integer of 1 to 20；

N is 0 or integer of 1 to 20；

The integer that the summation of (n+ [(h+j) k]+m) is 11 to 25；

When k is 1, then it is hydrogen that m, which is at least 2 and M,；And

When k is 0 and R is fluorine, then it is hydrogen that m, which is at least 2 and M,.

7. microfluidic device according to any one of claim 1 to 6, wherein the electrowetting of the equipment, which configures, includes The first part of the equipment, and wherein, the equipment further includes the second part that there is dielectrophoresis DEP to configure.

8. a kind of microfluidic device, including：

Substrate has at least one electrode for being configured to connect to voltage source；

Lid has at least one electrode for being configured to connect to the voltage source；And

At least one spacer element,

Wherein, the substrate and the lid are substantially parallel to each other, and are bonded together by the spacer element to limit quilt It is configured to keep the capsule of liquid,

Wherein, there are the substrate drop actuation surfaces for partly limiting the capsule, the drop actuation surfaces to have interior Dielectric layer and outer hydrophobic layer,

Wherein, the outer hydrophobic layer includes the self-association molecule on the surface for being covalently bound to the inner-dielectric-ayer, on it The hydrophobic monolayer of dense packing is formed, and

Wherein, when at least one electrode of at least one electrode of the substrate and the lid is connected to the voltage When the opposing terminal in source, the substrate being capable of pair aqueous drop contacted with the drop actuation surfaces of the substrate application electricity Wetting power.

9. microfluidic device according to claim 8, wherein the self-association molecule of the hydrophobic monolayer includes each surface Modified ligand and the surface modified ligand is linked to the inner-dielectric-ayer surface the link group, wherein the drop Actuation surfaces have the structure of Formula II：

Wherein,It is the surface of dielectric layer；

V is-P (O) (OY) W- or-Si (OZ)₂W-；

W is-O- ,-S- or-NH- and is connected to the surface；

R is hydrogen or fluorine；

M is hydrogen or fluorine；

H is independently 2 or 3 integer；

J is 1；

K is 0 or 1；

M is 0 or integer of 1 to 20；

N is 0 or integer of 1 to 20；

The integer that the summation of (n+ [(h+j) k]+m) is 11 to 25；

When k is 1, then it is hydrogen that m, which is at least 2 and M,；And

10. microfluidic device according to claim 9, wherein V is-Si (OZ)₂W-。

11. microfluidic device according to claim 9, wherein V is-P (O) (OY) W-.

12. the microfluidic device according to any one of claim 9 to 11, wherein n is integer of 1 to 20, and its In, R is hydrogen.

13. microfluidic device according to claim 12, wherein m is integer of 1 to 20, and wherein, and M is hydrogen.

14. microfluidic device according to claim 13, wherein m is 2.

15. the microfluidic device according to any one of claim 9 to 11, wherein n is integer of 1 to 20, and its In, R is fluorine.

16. microfluidic device according to claim 15, wherein m is integer of 1 to 20, and wherein, and M is hydrogen.

17. microfluidic device according to claim 16, wherein m is 2.

18. microfluidic device according to claim 9, wherein k is 1.

19. microfluidic device according to claim 9, wherein k is 0.

20. microfluidic device according to claim 9, wherein the summation of (n+ [(h+j) k]+m) be 13 to 19 it is whole Number.

21. microfluidic device according to claim 8, wherein the drop actuation surfaces of the substrate it is described outer Hydrophobic layer has the thickness less than 5 nanometers.

22. microfluidic device according to claim 8, wherein the drop actuation surfaces of the substrate it is described outer Hydrophobic layer is patterned such that selection area is relative hydropathic compared with the rest part of the outer hydrophobic layer.

23. microfluidic device according to claim 8, wherein the drop actuation surfaces of the substrate it is described in Dielectric layer includes oxidiferous first layer dielectric material.

24. microfluidic device according to claim 23, wherein the oxide is metal oxide.

25. microfluidic device according to claim 24, wherein the metal oxide is aluminium oxide.

26. microfluidic device according to claim 23, wherein the first layer dielectric material passes through atomic layer deposition shape At.

27. the microfluidic device according to any one of claim 23 to 26, wherein the drop of the substrate activates The inner-dielectric-ayer on surface further includes second layer dielectric material, and wherein, and the outer hydrophobic layer is covalently bound to described One layer of dielectric material.

28. microfluidic device according to claim 27, wherein the second layer dielectric material includes oxide or nitridation Object.

29. microfluidic device according to claim 28, wherein the second layer dielectric material be selected from by silica and The group that silicon nitride is constituted.

30. microfluidic device according to claim 27, wherein the second layer dielectric material passes through plasma enhancing Chemical vapor deposition is formed.

31. microfluidic device according to claim 23, wherein the first layer dielectric material includes the first sublayer dielectric Material and the second sublayer dielectric material, wherein first sublayer is covalently bound to the hydrophobic layer.

32. microfluidic device according to claim 31, wherein the first sublayer dielectric material includes silica.

33. microfluidic device according to claim 31, wherein the first sublayer dielectric material passes through ALD deposition.

34. the microfluidic device according to any one of claim 31 to 33, wherein the first layer dielectric material has The thickness of about 10nm to about 20nm.

35. microfluidic device according to claim 34, wherein the first sublayer dielectric material has about 2nm to about The thickness of 10nm.

36. microfluidic device according to claim 8, wherein the drop actuation surfaces of the substrate it is described in Dielectric layer has at least about 40 nanometers of thickness.

37. microfluidic device according to claim 36, wherein the drop actuation surfaces of the substrate it is described in Dielectric layer has about 40 nanometers to about 120 nanometers of thickness.

38. microfluidic device according to claim 8, wherein the substrate further includes photoresponsive layer, the photoresponsive layer The second side with the first side and contact at least one electrode that contact the inner-dielectric-ayer.

39. according to the microfluidic device described in claim 38, wherein the photoresponsive layer includes amorphous silicon hydride a-Si:H.

40. the microfluidic device according to claim 38 or 39, wherein the photoresponsive layer is at least 900 nanometers Thickness.

41. microfluidic device according to claim 40, wherein the photoresponsive layer is with about 900 to 1100 nanometers Thickness.

42. according to the microfluidic device described in claim 38, wherein the photoresponsive layer includes multiple conductors, each conductor At least one electrode of the substrate is connected to via phototransistor switch with capable of controlling.

43. microfluidic device according to claim 8, wherein the substrate includes being configured to connect to AC voltage sources Single electrode, the single electrode includes layer of ito.

44. microfluidic device according to claim 8, wherein the substrate includes being configured to connect to AC voltage sources Single electrode, the single electrode includes conductive silicon layer.

45. microfluidic device according to claim 8, wherein the substrate includes multiple electrodes, and each electrode is configured To be connected to one or more AC voltage sources.

46. microfluidic device according to claim 45, wherein each electrode in the multiple electrode can be via crystal Pipe switch is connected to one in one or more AC voltage sources.

47. microfluidic device according to claim 8, wherein the lid, which has, partly limits the inside of the capsule The inner surface on surface, the lid has internal layer and outer hydrophobic layer, wherein the outer hydrophobic layer of the lid includes covalent It is attached to the self-association molecule on the surface of the internal layer of the lid, to be formed on the hydrophobic monolayer of dense packing.

48. microfluidic device according to claim 47, wherein the self-association molecule of the hydrophobic monolayer of the lid each wraps It includes surface modified ligand and the surface modified ligand is linked to the link group on the surface of the internal layer of the lid, In, the inner surface of the lid has the structure of Formula II：

Wherein,It is the surface of dielectric layer；

V is-P (O) (OY) W- or-Si (OZ)₂W-；

W is-O- ,-S- or-NH- and is connected to the surface；

R is hydrogen or fluorine；

M is hydrogen or fluorine；

H is independently 2 or 3 integer；

J is 1；

K is 0 or 1；

M is 0 or integer of 1 to 20；

N is 0 or integer of 1 to 20；

The integer that the summation of (n+ [(h+j) k]+m) is 11 to 25；

When k is 1, then it is hydrogen that m, which is at least 2 and M,；And

49. microfluidic device according to claim 48, wherein the self-association molecule of the hydrophobic monolayer of the lid with it is described The self-association molecule of the hydrophobic monolayer of the drop actuation surfaces of substrate is identical.

50. the microfluidic device according to any one of claim 47 to 49, wherein the outer of the inner surface of the lid is dredged Water layer has the thickness less than 5 nanometers.

51. microfluidic device according to claim 47, wherein the internal layer of the lid is inner-dielectric-ayer.

52. microfluidic device according to claim 51, wherein the lid further includes photoresponsive layer.

53. microfluidic device according to claim 51, wherein the lid includes multiple electrodes, and each electrode is configured To be connected to one or more AC voltage sources.

54. microfluidic device according to claim 8, wherein at least one spacer element includes that silicon substrate is organic poly- Close object.

55. microfluidic device according to claim 54, wherein the silicon substrate organic polymer is selected from by poly dimethyl silicon The group that oxygen alkane PDMS and light patternable polysiloxanes PPS are constituted.

56. microfluidic device according to claim 8, wherein at least one spacer element includes SU-8.

57. microfluidic device according to claim 8, wherein at least one spacer element has at least 30 microns Thickness.

58. microfluidic device according to claim 8, wherein at least one spacer element limits in the capsule Fixed one or more microchannels.

59. microfluidic device according to claim 58, wherein at least one spacer element in the capsule into One step limits multiple chambers, wherein each chamber is from least one microchannel openings.

60. a kind of method of manufacture microfluidic device, the method includes：

Spacer element is attached to the inner surface of the lid at least one electrode for being configured to connect to voltage source；

The spacer element and lid are attached to Jie of the substrate at least one electrode for being configured to connect to voltage source Ammeter face, thus the spacer element become to be arranged between the inner surface and the dielectric surface of the substrate of the lid, it is described Lid and the substrate are oriented substantially parallel to one another, and the substrate, spacer element and common limit of lid are configured to keep The capsule of liquid；

The hydrophobic monolayer of dense packing is formed at least part of the inner surface of the lid by being vapor-deposited, wherein institute State the self-association molecule that hydrophobic monolayer includes the inner surface for being covalently bound to the lid；And

The hydrophobic monolayer of dense packing is formed at least part of the dielectric surface of the substrate by being vapor-deposited, In, the hydrophobic monolayer includes the self-association molecule for the dielectric surface for being covalently bound to the substrate.

61. method according to claim 60, wherein the self-association molecule of the hydrophobic monolayer of the lid and the substrate The self-association molecule of hydrophobic monolayer includes each surface modified ligand and the surface modified ligand is linked to the lid respectively Inner surface and the substrate dielectric surface the link group, wherein it is described lid and the substrate gained surface have formula The structure of II：

Wherein,It is the surface of dielectric layer；

V is-P (O) (OY) W- or-Si (OZ)₂W-；

W is-O- ,-S- or-NH- and is connected to the surface；

R is hydrogen or fluorine；

M is hydrogen or fluorine；

H is independently 2 or 3 integer；

J is 1；

K is 0 or 1；

M is 0 or integer of 1 to 20；

N is 0 or integer of 1 to 20；

The integer that the summation of (n+ [(h+j) k]+m) is 11 to 25；

When k is 1, then it is hydrogen that m, which is at least 2 and M,；And

62. method according to claim 61, wherein V is-Si (OZ)₂W-。

63. method according to claim 61, wherein V is-P (O) (OY) W-.

64. method according to claim 61, wherein n is integer of 1 to 20, and wherein, and R is hydrogen.

65. method according to claim 64, wherein m is integer of 1 to 20, and wherein, and M is hydrogen.

66. method according to claim 65, wherein m is 2.

67. method according to claim 61, wherein n is integer of 1 to 20, and wherein, and R is fluorine.

68. method according to claim 67, wherein m is integer of 1 to 20, and wherein, and M is hydrogen.

69. method according to claim 68, wherein m is 2.

70. the method according to any one of claim 61 to 69, wherein k is 1.

71. the method according to any one of claim 61 to 69, wherein k is 0.

72. microfluidic device according to claim 61, wherein the summation of (n+ [(h+j) k]+m) be 13 to 19 it is whole Number.

73. a kind of microfluidic device, including：

Conductive silicon substrate, with dielectric stack and at least one electrode for being configured to connect to voltage source；

Lid has at least one electrode for being configured to connect to voltage source；And

At least one spacer element,

Wherein, the conductive silicon substrate and it is described lid be substantially parallel to each other and by the spacer element be bonded together with The capsule for being configured as keeping liquid is limited,

Wherein, the conductive silicon substrate has the inner surface for partly limiting the capsule, described inwardly facing surface including described The outmost surface of dielectric stack, and

Wherein, when at least one electrode of at least one electrode of the substrate and the lid is connected to AC voltage sources Opposing terminal when, the substrate can pair aqueous drop contacted with the inner surface of the substrate apply electrowetting power.

74. according to the microfluidic device described in claim 73, wherein the conductive silicon substrate includes non-crystalline silicon.

75. according to the microfluidic device described in claim 73, wherein the conductive silicon substrate includes photo-transistor arrays.

76. according to the microfluidic device described in claim 73, wherein the conductive silicon substrate includes electrod-array.

77. according to the microfluidic device described in claim 73, wherein the inner surface of the conductive silicon substrate further includes outer dredges Water layer, the outer hydrophobic layer include the self-association molecule for being covalently bound to the interior dielectric stack.

78. according to the microfluidic device described in claim 73, wherein the interior dielectric stack includes first layer dielectric material With second layer dielectric material.

79. according to the microfluidic device described in claim 78, wherein the first layer dielectric material has first surface and phase To surface, wherein the first surface of the first layer abuts the second layer, and wherein, the apparent surface of the first layer Form the outmost surface of the dielectric stack.

80. according to the microfluidic device described in claim 78, wherein the first layer dielectric material includes metal oxide.

81. according to the microfluidic device described in claim 80, wherein the first layer dielectric material includes aluminium oxide or oxidation Hafnium.

82. according to the microfluidic device described in claim 78, wherein the second layer dielectric material includes oxide or nitridation Object.

83. according to the microfluidic device described in claim 82, wherein the second layer dielectric material includes silica or nitridation Silicon.

84. according to the microfluidic device described in claim 78, wherein the second layer passes through plasma enhanced chemical vapor PECVD technique is deposited to deposit.

85. according to the microfluidic device described in claim 78, wherein the first layer is heavy by atomic layer deposition ALD technique Product.

86. according to the microfluidic device described in claim 78, wherein the interior dielectric stack include have first surface and The third layer of apparent surface, wherein the first surface of the third layer abuts the apparent surface of the first layer, and wherein, The apparent surface of the third layer forms the outmost surface of the dielectric stack.

87. according to the microfluidic device described in claim 86, wherein the third layer includes silica.

88. according to the microfluidic device described in claim 86, wherein the third layer is heavy by atomic layer deposition ALD technique Product.

89. according to the microfluidic device described in claim 78, wherein the first layer dielectric material has about 10nm to about The thickness of 50nm.

90. according to the microfluidic device described in claim 86, wherein the first layer dielectric material has about 5nm to about The thickness of 20nm and the third layer dielectric material have the thickness of about 2nm to about 10nm.

91. according to the microfluidic device described in claim 78, wherein the second layer dielectric material has about 30nm to about The thickness of 100nm.

92. according to the microfluidic device described in claim 73, wherein the dielectric stack of the drop actuation surfaces of the substrate With at least about 40 nanometers of thickness.

93. according to the microfluidic device described in claim 92, wherein the dielectric stack of the drop actuation surfaces of the substrate With about 40 nanometers to about 120 nanometers of thickness.

94. according to the microfluidic device described in claim 73, wherein the dielectric layer is with about 50k ohm to the Europe about 150k The impedance of nurse.

95. the microfluidic device according to any one of claim 73 to 94, wherein described device includes：

Dielectrophoresis module, for executing the first microfluidic procedures in response to the first application voltage under first frequency；And

Electrowetting module for receiving the output from the dielectrophoresis module, and is applied in response to second under second frequency Making alive executes the second microfluidic procedures,

Wherein, the Electrowetting module includes the dielectric stack of the conductive silicon substrate.

Further include the bridge between the first module and the second module 96. according to the microfluidic device described in claim 95.

97. according to the microfluidic device described in claim 96, wherein the bridge does not execute first microfluidic procedures or institute State the second microfluidic procedures.

98. according to the microfluidic device described in claim 96, wherein the bridge is charge neutrality.

99. according to the microfluidic device described in claim 96, wherein the bridge includes pipeline.

100. according to the microfluidic device described in claim 96, wherein the bridge includes polymer.

101. according to the microfluidic device described in claim 95, wherein the output is biomaterial.

102. according to the microfluidic device described in claim 95, wherein range of the first frequency in 100kHz to 10mHz It is interior.

103. according to the microfluidic device described in claim 95, wherein range of the second frequency in 1kHz to 300kHz It is interior.

104. according to the microfluidic device described in claim 95, wherein the first voltage is in the range of 1 to 10 volt.

105. according to the microfluidic device described in claim 95, wherein the second voltage is in the range of 10 to 100 volt.

106. according to the microfluidic device described in claim 95, wherein the conductive silicon substrate is whole.

107. according to the microfluidic device described in claim 95, wherein the conductive silicon substrate is binary.

108. according to the microfluidic device described in claim 106, wherein the conductive silicon substrate includes non-crystalline silicon.

109. according to the microfluidic device described in claim 107, wherein the conductive silicon substrate includes non-crystalline silicon.

110. according to the microfluidic device described in claim 106, wherein the conductive silicon substrate includes phototransistor battle array Row.

111. according to the microfluidic device described in claim 107, wherein the conductive silicon substrate includes phototransistor battle array Row.

112. according to the microfluidic device described in claim 106, wherein the conductive silicon substrate includes electrod-array.

113. according to the microfluidic device described in claim 107, wherein the conductive silicon substrate includes electrod-array.

114. it is a kind of be used for transmission speck body that is compatible with aqueous medium and/or being dissolved in aqueous medium, biological product and/or The system of reagent, the system comprises：

Microfluidic device, with the capsule for including base portion and microfluidic circuit structure, wherein the base portion includes being covalently bound to At least part of hydrophobic monolayer of the upper surface of the base portion；

The first fluid medium unmixing with aqueous medium；And

At least one aqueous drop.

115. according to the system described in claim 114, wherein the hydrophobic monolayer has surface modified ligand and changes surface Property ligand link to the link group on the surface, wherein the hydrophobic surface has the structure of Formula II：

Wherein,It is the surface；

V is-P (O) (OY) W- or-Si (OZ) 2W-；

W is-O- ,-S- or-NH- and is connected to the surface；

R is hydrogen or fluorine；

M is hydrogen or fluorine；

H is independently 2 or 3 integer；

J is 1；

K is 0 or 1；

M is 0 or integer of 1 to 20；

N is 0 or integer of 1 to 20；

The integer that the summation of (n+ [(h+j) k]+m) is 11 to 25；

When k is 1, then it is hydrogen that m, which is at least 2 and M,；And

116. according to the system described in claim 114, wherein the base portion includes conductive substrates.

117. according to the system described in claim 114, wherein the microfluidic device is appointed according in claim 1 to 59 Microfluidic device described in one.

118. according to the system described in claim 117, wherein the microfluidic device includes the EW configurations of optical actuation.

119. according to the system described in claim 117, wherein the microfluidic device further includes DEP configurations.

120. according to the system described in claim 114, wherein the first fluid medium includes having backbone structure at least A kind of organic compound or at least one organo-silicon compound, the backbone structure include the atom selected from carbon, silicon and oxygen.

121. according to the system described in claim 120, wherein the backbone structure of at least one organo-silicon compound includes Silicon atom and optional oxygen atom.

122. according to the system described in claim 120, wherein the backbone structure of at least one organic compound includes carbon Atom and optional oxygen atom.

123. according to the system described in claim 122, wherein the backbone structure is branch.

124. according to the system described in claim 120, wherein the first fluid medium, which includes that one or more are acyclic, to be had Machine or organo-silicon compound.

125. according to the system described in claim 124, wherein the first fluid medium is by acyclic organic or organosilicon compound Object forms.

126. according to the system described in claim 114, wherein the first fluid medium does not include perfluocarbon atom.

127. according to the system described in claim 114, wherein the substitution of the carbon atom of the compound of the first fluid medium Base includes the fluoro substituents no more than 90%.

128. according to the system described in claim 115, wherein the surface modified ligand includes at least first part, described First part includes perfluocarbon atom in the end towards inside of the hydrophobic monolayer.

129. according to the system described in claim 128, wherein whole carbon atoms of the hydrophobic monolayer are fluoridized.

130. according to the system described in claim 114, wherein the first fluid medium includes more than one organic or organic Silicon compound.

131. according to the system described in claim 114, wherein the capsule further includes lid.

132. according to the system described in claim 131, wherein the lid is transparent to light.

133. according to the system described in claim 131, wherein the lid includes glass and/or indium oxide tantalum ITO.

134. the system according to any one of claim 131 to 133, wherein the lid includes electrode.

135. according to the system described in claim 114, wherein the aqueous drop includes surfactant.

136. according to the system described in claim 135, wherein the surfactant includes nonionic surfactant.

137. the system according to claim 135 or 136, wherein the surfactant includes the copolymerization of block oxyalkylene Object, aliphatic ester ethoxylation dehydrated sorbitol, ethoxylation fluorine-containing surfactant, lauryl sodium sulfate or 2,4,7, 9- tetramethyl -5- decine -4,7- diol ethoxylates.

138. according to the system described in claim 135, wherein the surfactant includes FS-30 (DuPontTM, Synquest Laboratories).

139. according to the system described in claim 114, wherein the drop includes phosphate buffered saline.

140. according to the system described in claim 114, wherein the aqueous drop includes at least one speck body.

141. according to the system described in claim 140, wherein the speck body is biological speck body.

142. according to the system described in claim 114, wherein the aqueous drop includes containing nucleic acid and/or protein Biological product.

143. according to the system described in claim 114, wherein the aqueous drop includes reagent.

144. a kind of being used for transmission speck body that is compatible with aqueous medium and/or dissolving in aqueous medium, biological product and/or examination The kit of agent, the kit include：

Microfluidic device, with the capsule for including base portion and microfluidic circuit structure, wherein the base portion includes being covalently bound to At least part of hydrophobic monolayer of the upper surface of the base portion；And

The first fluid medium unmixing with aqueous medium.

145. according to the kit described in claim 144, wherein hydrophobic monolayer has surface modified ligand and by the surface Modified ligand links to the link group on the surface, wherein the hydrophobic surface has the structure of Formula II：

Wherein,It is surface；

V is-P (O) (OY) W- or-Si (OZ) 2W-；

W is-O- ,-S- or-NH- and is connected to the surface；

R is hydrogen or fluorine；

M is hydrogen or fluorine；

H is independently 2 or 3 integer；

J is 1；

K is 0 or 1；

M is 0 or integer of 1 to 20；

N is 0 or integer of 1 to 20；

The integer that the summation of (n+ [(h+j) k]+m) is 11 to 25；

When k is 1, then it is hydrogen that m, which is at least 2 and M,；And

146. according to the kit described in claim 144, wherein the base portion includes conductive substrates.

147. kit according to any one of claim 144 to 146, wherein the microfluidic device is according to power Profit requires the microfluidic device described in any one of 1 to 72.

A kind of 148. processes operating the microfluidic device according to any one of claim 8 to 59, the process include：

A part for the capsule or the capsule is filled with the first liquid medium；

Apply AC voltage potentials between at least one electrode of the substrate and at least one electrode of the lid；

First drop of liquid is introduced into the capsule, wherein first drop is immiscible in first liquid medium In；And

By applying electrowetting power, the desired locations first drop being moved in the capsule to first drop.

149. according to the process described in claim 148, wherein first liquid medium is oil.

150. according to the process described in claim 148, wherein first liquid medium is silicone oil, fluorinated oil or combinations thereof.

151. according to the process described in claim 148, wherein the AC voltage potentials applied are at least 20ppV.

152. according to the process described in claim 151, wherein the AC voltage potentials applied are between about 25 and 35ppV.

153. according to the process described in claim 148, wherein the AC voltage potentials applied have about 1 to 100kHz frequency Rate.

154. according to the process described in claim 148, wherein and the microfluidic device includes droplet generator, and wherein, First drop is introduced the capsule by the droplet generator.

155. according to the process described in claim 148, wherein the first drop packet aqueous solution.

156. according to the process described in claim 155, wherein first drop includes at least one speck body.

157. according to the process described in claim 156, wherein at least one speck body is biological speck body.

158. according to the process described in claim 157, wherein the biology speck body is cell.

159. according to the process described in claim 155, wherein the aqueous solution is cell culture medium.

160. according to the process described in claim 156, wherein at least one speck body is that have to interested material The capture bead of affinity.

161. according to the process described in claim 160, wherein first drop includes 2 to 20 capture beads.

162. according to the process described in claim 160, wherein the interested material is biological cell secretion.

163. according to the process described in claim 160, wherein the interested material is selected from by DNA, genomic DNA, line Mitochondrial DNA, RNA, mRNA, miRNA or any combination thereof composition group.

164. according to the process described in claim 155, wherein first drop includes reagent.

165. according to the process described in claim 164, wherein the reagent is cell cracking agent.

166. according to the process described in claim 165, wherein the reagent includes non-ionic detergent.

167. according to the process described in claim 166, wherein the concentration of the non-ionic detergent is less than 0.2%.

168. according to the process described in claim 164, wherein the reagent is proteolytic enzyme.

169. according to the process described in claim 168, wherein the proteolytic enzyme can be deactivated.

170. according to the process described in claim 148, further includes：

Second drop of liquid is introduced into the capsule, wherein the liquid of second drop is in first liquid medium In it is unmixing but miscible with the liquid of first drop；

By the way that electrowetting power is applied to second drop, second drop is moved to adjacent with first drop Position in the capsule；And

Merge second drop and combines drop with first drop to form first.

171. according to the process described in claim 170, wherein by being applied to second drop and/or first drop Power-up wetting power makes second drop and first droplet coalescence.

172. according to the process described in claim 170, wherein first drop includes biological speck body, and wherein, institute It includes reagent to state the second drop.

173. according to the process described in claim 172, wherein the reagent for including in second drop is selected to be buffered by cracking The group that liquid, fluorescent marker and luminescence assays reagent are constituted.

174. according to the process described in claim 172, wherein and the reagent for including in second drop is lysis buffer, And wherein, the biological cell is cleaved in first drop and second droplet coalescence.

175. according to the process described in claim 170, further includes：

The third drop of liquid is introduced into the capsule, wherein the liquid of the third drop is in first liquid medium In liquid unmixing but that drop is combined with described first it is miscible；And

By the way that electrowetting power is applied to the third drop, the third drop is moved in the capsule and described first Combine the adjacent position of drop；And

176. according to the process described in claim 175, wherein by the third drop and/or the first combination liquid Drop, which applies electrowetting power, makes the third drop combine droplet coalescence with described first.

177. according to the process described in claim 175, wherein the third drop includes reagent.

178. according to the process described in claim 177, wherein the third drop includes protease inhibitors.

179. according to the process described in claim 177, wherein the third drop includes to interested material with affine One to 20 capture bead of power.

180. according to the process described in claim 179, wherein capture bead includes oligonucleotides capturing agent.

181. according to the process described in claim 180, wherein the oligonucleotides capturing agent is poly- dT oligonucleotides.

182. according to the process described in claim 179, wherein the interested material is selected from by DNA, genomic DNA, line Mitochondrial DNA, RNA, mRNA, miRNA or any combination thereof constitute group.

183. according to the process described in claim 179, further includes：

From microfluidic device output one to 20 capture bead.

184. process according to according to claim 175 further includes：

4th drop of liquid is introduced into the capsule, wherein the liquid of the 4th drop is in first liquid medium In liquid unmixing but that drop is combined with described second it is miscible；

By the way that electrowetting power is applied to the 4th drop, the 4th drop is moved to and combines drop phase with described second Position in the adjacent capsule；And

185. according to the process described in claim 184, wherein by the 4th drop and/or the second combination liquid Drop, which applies electrowetting power, makes the 4th drop combine droplet coalescence with described second.

186. according to the process described in claim 184, wherein the 4th drop includes reagent.

187. according to the process described in claim 186, wherein the reagent for including in the 4th drop includes containing buffering Liquid, dNTP and be adapted for carrying out reverse transcription reaction polymerase mixture.

188. according to the process described in claim 186, wherein the reagent for including in the 4th drop includes containing buffering Liquid, dNTP and the mixture for being adapted for the polymerase that whole genome amplification reacts.

189. according to the process described in claim 148, wherein first drop, the second drop, third drop and the 4th liquid Drop is respectively with about 5 to 50 nanoliters of volume.

190. according to the process described in claim 189, wherein first drop, second drop and the third liquid Drop is respectively with about 5 to 20 nanoliters of volume.

191. according to the process described in claim 190, wherein the volume base of second drop and/or the third drop It is equal to the volume of first drop in sheet.

192. according to the process described in claim 190, wherein it is larger about 1 that the 4th drop, which has than first drop, To 3 times of volume.

193. according to the process described in claim 192, wherein the 4th drop has about 10 to 30 nanoliters of volume.

194. according to the process described in claim 148, wherein the capsule includes at least one microchannel.

195. according to the process described in claim 194, wherein first drop is moved to the expectation position in the capsule It sets including first drop is moved through at least one microchannel.

196. according to the process described in claim 194, wherein the capsule further includes from least one microchannel openings Multiple chambers.

197. according to the process described in claim 196, wherein first drop is moved to the expectation position in the capsule It sets including first drop to be moved in the multiple chamber chamber.

198. according to the process described in claim 194, wherein is moved to second drop adjacent with first drop Position include that second drop is moved through at least one microchannel and is optionally moved to comprising described In the chamber of one drop.

199. according to the process described in claim 198, wherein is moved to the third drop and combines drop with described first Adjacent position includes that the third drop is moved through at least one microchannel and is optionally moved to comprising institute In the chamber for stating the first combination drop.

200. according to the process described in claim 199, wherein is moved to the 4th drop and combines drop with described second Adjacent position includes that the 4th drop is moved through at least one microchannel and is optionally into comprising described In the chamber of second combination drop.

201. process according to according to claim 148, wherein apply electrowetting power with mobile and/or merging drop packet Include effective electro-wetting property in the region close to drop for changing the substrate surface.

202. according to the process described in claim 201, wherein it includes activating the substrate surface to change effective electro-wetting property The region close to drop at electrowetting electrode.

203. according to the process described in claim 202, wherein the substrate includes photoresponsive layer, and wherein, described in activation Electrowetting electrode at the region close to drop of substrate surface includes that light pattern is directed to described in the electrowetting surface On region.