CN106104271B - Micro-fluid chip and its manufacture with conical bead trapping chamber - Google Patents
Micro-fluid chip and its manufacture with conical bead trapping chamber Download PDFInfo
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- CN106104271B CN106104271B CN201580012689.2A CN201580012689A CN106104271B CN 106104271 B CN106104271 B CN 106104271B CN 201580012689 A CN201580012689 A CN 201580012689A CN 106104271 B CN106104271 B CN 106104271B
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
- B01L2200/0668—Trapping microscopic beads
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/0864—Configuration of multiple channels and/or chambers in a single devices comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/0867—Multiple inlets and one sample wells, e.g. mixing, dilution
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0887—Laminated structure
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/08—Regulating or influencing the flow resistance
- B01L2400/084—Passive control of flow resistance
- B01L2400/086—Passive control of flow resistance using baffles or other fixed flow obstructions
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/554—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being a biological cell or cell fragment, e.g. bacteria, yeast cells
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01L—SEMICONDUCTOR DEVICES NOT COVERED BY CLASS H10
- H01L21/00—Processes or apparatus adapted for the manufacture or treatment of semiconductor or solid state devices or of parts thereof
Abstract
A kind of micro-fluid chip (100), including layer (10,60), the array (30) of pearl trapping chamber (20) is located in the layer (10,60), each of wherein described chamber (20) has the taper limited by one or more side walls (21 24), the side wall is hydrophilic, and each in wherein described chamber (20) extends as blind hole in the thickness of the layer (10,60).The manufacture method of chip as also providing.
Description
Technical field
The present invention relates generally to the field of micro-fluid chip, and particularly for example for bioanalysis application
, micro-fluid chip equipped with pearl trapping chamber, and relative manufacturing process.
Background technology
Microfluidic device is generally referred to as micro manufacturing device, it is used to pump, sample, mix, analyze and quantify to liquor charging
Body.The prominent features of microfluidic device are derived from the special behavior that liquid shows on micrometer length scales.Liquid in microfluidic device
The flowing of body can be typically stratiform.Volume far below one nanoliter can be by manufacturing with the lateral ruler in micrometer range
Very little structure obtains.Therefore the reaction for (passing through the diffusion of reactant) in large scale and being restricted can be accelerated.Microfluid
Device thus be accordingly used in various applications.
Many microfluidic devices have user's chip interface and closure flow path.Flow path is closed to be easy to Functional Unit
Part (for example, heater, mixer, pump, UV detector, valve etc.) is incorporated into a device, at the same minimize with leakage and
The problem of evaporation is related.
In general, the acceptor on surface is used to combine needs detected specific analyte in the sample.After the coupling, sample
Product and interfering material can be washed off.Then can be directly (such as change via quality, refractive index etc.) or indirectly (glimmering
Light immunoassays etc.) detection acceptor-analyte complex.Although microfluidic device is the promising device for analysis,
Be acceptor is incorporated at present micro-fluid chip inside it is very challenging.
Solution is had pointed out, wherein acceptor is patterned on the surface of micro-fluid chip.In more detail, microfluid core
Piece can be sealed with the PDMS layer for the line for being patterned with capture antibody thereon.In this case, by using masterplate from molten
Liquid absorption antibody realizes patterning capture antibody.However, such method is troublesome, there is poor efficiency, it is necessary to masterplate, and
It is and additionally, due to adsorption process and slower.In addition, PDMS is expensive and contaminated surface is (after~20 minute contact times
Make the water-wetted surface hydrophobic).
Other solutions known for a long time rely on the microballon for measuring.Herein, pearl be typically coated with by
Body.Pearl (for example, magnetic bead, or single pearl in capillary) or can deposit/be positioned at the spy of micro-fluid chip in the solution
Used after determining in region.Two kinds of situations can be distinguished：
Magnetic bead：Pearl is separated from interfering material and sample with flushing by using magnet.But magnetic bead is than non-magnetic bead more
It is expensive and be difficult to prepare.Moreover, these pearls are opaque and not are suitable for very much optics/fluorescence-based measure；With
And
Non- magnetic bead, develops many technologies for positioning/manipulation pearl in microfluidic device for non-magnetic bead.
However, such technology has following defect.They are needed：
- particular activated device (electrode, magnetic texure, focuses on light, transducer, piezoelectric structure etc.), and it is therefore complicated and
It is expensive；Or
The geometry in particular (certain radius of curvature, contraction flow region) in-microfluidic flow path, it should be noted that with and without
Pearl, the flowed friction of chip can be significantly different, and the constancy system for the pearl being trapped it is problematic.It is moreover, specific
The viscosity of sample/liquid can be problem.
As described above, some solutions use contraction flow region or " filter ", it is directly the flowing in micro-fluid chip
The part in path, for trapping pearl.However, such solution causes the balance between signal strength and signal quality.
Pearl stability is further problematic.
The present invention proposes the definite solution for pearl stability problem.
The content of the invention
According in a first aspect, the present invention is embodied as a kind of micro-fluid chip, it includes layer, and the array of pearl trapping chamber is set
In said layer, wherein each in the chamber has the taper limited by one or more side walls, the side wall is parent
Water, and each of wherein described chamber extends as blind hole in the thickness of the layer.
In embodiment, at least some with the pyramid shape formed by side wall in the chamber, the side wall is parent
Water, and wherein it is preferred to, which is substantially limited by least four sidewalls.
Preferably, the layer includes one or more semiconductor elements of such as silicon, and the pyramid shape tool of the chamber
There is the geometry consistent with the anisotropic etching process of the manufacture of the chamber in the layer.
In a preferred embodiment, at least one, the preferably major part in the chamber are filled with pearl, preferably microballon, it is flat
It is respectively provided between 1 to 40 μm, the diameter preferably between 2 to 20 μm and more preferably between 2 to 10 μm.
Preferably, the major part in the chamber of the array is filled with only one pearl, it preferably averagely has 1 to arrive
The microballon of diameter between 40 μm, preferably between 2 to 20 μm and more preferably between 2 to 10 μm.
In embodiment, the ratio of the average-size of the opening of the chamber and the average diameter of the pearl in the chamber is 1.0
To between 2.4, and preferably between 1.4 to 2.0.
Preferably, the ratio of the mean depth of the chamber and the average diameter of the pearl in the chamber is at least 0.5, preferably
1.0, and more preferably 1.3.
In a preferred embodiment, two or more at least one subgroups in the chamber are connected by least one microchannel
Connect, the subgroup is preferably the row or column of the array of the chamber, and wherein, it is highly preferred that one in the subgroup or
Multiple to be defined in channel portion, the bottom wall or roof of the channel portion are formed by the surface of the layer.
Preferably, the array is sealed by the cap rock of the array extension relative to the chamber.
In embodiment, the chip includes some arrays of one or more pearl trapping chambers, wherein the array is preferred
Ground is inserted between different pairs of channel portion.
Preferably, the chip includes being correspondingly situated at least two in some arrays one or more chambers
At least two different type pearls, wherein the different types of pearl is preferably in terms of size, coating, material and/or color
It is different.
According on the other hand, the present invention is embodied as a kind of any one microfluid core in above example
The manufacture method of piece, the described method includes：
Micro-fluid chip main body with layer is provided；And
The array of pearl trapping chamber is manufactured in said layer, wherein each in the chamber has by one or more side walls
The taper of restriction, the side wall be it is hydrophilic, and each of wherein described chamber as blind hole in the thickness of the layer
Extension.
Preferably, the anisotropic etching process that manufacturing the array includes being preferably used from limit is etched anisotropically through
The layer is to obtain the chamber.
In embodiment, the method further includes：The institute of the array is deposited to by spreading the drop of pearl solution by pearl
State in chamber；And the array is sealed with the cap rock that the array being positioned to relative to the chamber extends, and wherein described in sealing
Array preferably includes the lamination cap rock.
Preferably, after pearl is deposited and before sealing, the method further includes in the following order：Drying is at it
In have pearl the chamber array；Preferably the array is rinsed by using rinse solution and/or be used to adhere to by applying
The band of excessive pearl removes the excessive pearl for the inside for not being trapped in the chamber；And if necessary, it is dry again that there is pearl wherein
The array of the chamber.
By non-limitative example and specific implementation apparatus and method of the present invention will be described with reference to the drawings now.In figure
Shown technical characteristic is not necessarily drawn to.
Brief description of the drawings
Fig. 1, Fig. 2, Figure 12 and Figure 13 show that simplifying for the micro-fluid chip of different embodiment according to the subject invention represents
2D (bow) view；
Fig. 3 is shown：The pearl being trapped in pyramid chamber being related in embodiment simplifies the sectional view (above) represented
With top view (figure below)；
Fig. 4 is the scanning electron microscopy picture for the pearl being trapped in pyramid chamber being related in embodiment；
Fig. 5 A-5I schematically show the detailed step of the manufacture method of micro-fluid chip according to an embodiment of the invention
Suddenly；
Fig. 6 schematically shows the selected step of the manufacture method of Fig. 5, wherein upper row shows the section of micro-fluid chip
Figure, and lower row shows corresponding top view；
Similarly, Fig. 7-9 schematically shows the selected step of the modification of the manufacture method of Fig. 5；
Figure 10 corresponds to the flow chart of Fig. 5；
Figure 11 is shown in the 2D views for simplifying expression of the improved pyramid chamber involved in embodiment.Leftmost diagram is to bow
View, and other figures are sectional views；
Figure 14 shows the egative film of the fluoroscopic image for the pearl being trapped in the array of the chamber obtained according to an embodiment of the invention；
And
Figure 15 is the sectional view for simplifying expression for the pearl being trapped in frustoconical chamber being related in embodiment.
Embodiment
First, and generally with reference to Fig. 1-4 and 11-13, description it is related to the aspect of the invention of micro-fluid chip 100.
Chip significantly includes working lining 10,60, and the layer includes the array 30 for being located at pearl trapping chamber 20 therein (also referred to as trap).
It is apparent that each in chamber 20 has the taper surrounded by one or more side wall 21-24.Importantly, this or these side
Wall be hydrophilic and chamber 20 in each extend as blind hole in the thickness of working lining 10,60.
Working lining 10,60 can be substrate layer 10 or cap rock 60.Chamber is preferably disposed in the thickness of layer, the surface of the layer
It is microfluidic channel or the bottom wall or roof of channel part 12a, as shown in Figure 2.
In this application, " taper " also illustrates that " taper ", that is, has the form of similar cone.By " taper ", and with
One of cone is general to define as one man (http://en.wikipedia.org/wiki/Cone), it is represented from planar base portion
The opening 28 of chamber (correspond to) smoothly taper to the area of section smaller than base portion 28 opposed end (for example, vertex or
Truncation surface) 3 dimensional coil geometry, the planar base portion is not necessarily circle.Other definition assume that cone is the specific feelings of pyramid
Cone, i.e., is defined as the pyramid with circular cross section, for example, see http by condition://mathworld.wolfram.com/
Cone.html.It is assumed herein that taper includes the pyramid (having polygonal base) as particular case.It is that is, current
Taper need not have circular cross section.
Since manufacture limits, taper however will likely be truncated, or do not terminate as perfect, punctiform vertex at least.In fact,
Frustum even can be utilized, as described later.As being further illustrated in described immediately above and such as the following examples,
Bellmouth will preferably have polygonal base (base portion corresponds to the perforate of chamber 20), and therefore (have polygon for pyramid
Shape base portion).Taper can be referred to as bevel taper shape in this case, the reason is that pearl 50 (may pass through from direct distribution
Liquid dissemination technology) drop on the top of the lower array 30 of pearl trap is captured, and wherein trap is chamber 20, it is at top
Unlimited, i.e., inside village hollowing.However, the configuration can overturn, so that with the chamber being located in the cover, referring to Fig. 7.
The other solutions also tested compared to the present inventor, such as the side for trap using column or through hole shape chamber
Case, uses blind hole shape conical cavity result to prove unexpectedly favourable in terms of the stability of pearl for pearl trap.It is, by
This trap produced shows the unprecedented tendency for keeping pearl 50 after being flushed.The embodiment of the present invention is applied to microballon
(for example, there is the pearl of acceptor in its surface, wherein according to the Normal practice in bioassay, such acceptor can be used for
Ligand in binding soln), there is the complexity of appropriateness and be not required additional means (such as vacuum, magnetic field or electric field) to protect pearl
Hold in place.
The cause significantly improved of the stability of pearl is not clear；Improved stability is probably due to when pearl is trapped in chamber
In, i.e., at the tip of bellmouth when, laterally (conical cavity preferably protects pearl from fluid (gas or liquid) when rinsing
Lateral, move in plane influence) and/or the interface occurred below pearl and/or mechanical phenomenon.Moreover, (the one of conical cavity
It is a or multiple) side wall need be it is hydrophilic, in order to it work.
Current design and corresponding manufacture method also allow to place pearl before trap is covered, with majority (even if not being
Solution is different known to all).
In manufacture view, the present inventor is had been noted that after scattered tear drop solution and before excessive pearl is rinsed, to pearl
50 and trap 20 stability of the still substantial pearl for improving and being trapped is dried.This will then be described in detail.
Some known prior art solutions (for example, see, Sohn et al., biology sensor and bioelectronics device
Part, volume 21,2005, the 303-312 pages) in, through hole shape pyramid chamber is manufactured, so as to trapping " huge " pearl, i.e., straight
The pearl of magnitude more at least one greatly than pearl contemplated herein on footpath.However, since the pyramid chamber of manufacture is logical in this case
Hole, they necessarily have minimum-depth much too big for the application.Moreover, such solution need pickup-
The application of place tool and dorsal part vacuum, it may not apply to the pearl of smaller, microballon as contemplated herein, and microballon typically has
There are the polymeric beads of the diameter in the range of 1-40 μm (such pearl typically obtains from liquid suspension).
Although the present invention allows to obtain stable pearl in chamber 20, but it is not precluded from the additional of dielectric DEP electrodes etc.
Use.DEP electrodes can be physically located in trap or near, such as float pearl 50, so as to for example recycle pearl (if desired
If), or the solution that pearl is more exposed to flow in the microchannel residing for the array 30 in trap.In addition, in embodiment
In, the surface of chamber can for example be metallized using Al amplifies fluorescence signal will pass through reflection.
Pyramid can be endowed by referring more especially to some or all of Fig. 3, Fig. 4, Figure 11 and Figure 15, chamber 20 now
Shape, i.e., the taper volume with polygonal base, it is, the chamber formed by some, different lateral 21-24, the wall is being worked as
Before in the case of be hydrophilic.Pyramid shape for example can be tetrahedron, quadrangular pyramid or more complicated.Although preferably the shape
Substantially limited by (at least) four side wall 21-24.Therefore, such chamber 20 typically has triangular wall 21-24, it is preferable
Ground converges to a little, but is more likely to converge to truncation surface 29.The planar base portion for limiting the opening 28 of chamber has polygon, it is at least
It is three sides, and preferably four sides.In the latter case, pyramid shape is substantially by least four sidewalls 21-24
Limit.
It should be noted that wall needs not be perfect flat wall, it may be possible to due to the danger in manufacturing process.However, it should note
Meaning, preferably fabricated technology (see below) disclosed herein substantially prevent the situation.Such as due to photoetching process, some are residual
Surplus and deficiency falls into that there may be but this is actually rare.On the contrary, side wall is typically flat, the reason is that anisotropy is lost
Quarter follows crystal face.Such as can be by changing the shape of mask (layout) control as shown in Figure 11, so final form is at least
Can be controllable and predictable in embodiment.One or more of side wall for example can be structured or shape, such as
Shown in Figure 11, one in its mesospore is structured 23a-d, so that allow localized liquid to inject, and pearl release and attraction,
For those embodiments without using cap rock.
Compared to the chamber of " more " cone, in terms of the stability of pearl, it has been found that it is better to be caused using pyramid shape
As a result.So the reason for, is not entirely understood also.This is probably the free space due to being left in the turning around spherical bead,
This allows residual liquid to evaporate, different from cone shape referring to Fig. 3.
Fig. 4 is the scanning electron microscopy picture for showing the pearl being trapped in pyramid chamber.In the images：
- " EHT " represents the electronic high voltage in terms of kilovolt kV；
- " WD " represents the operating distance between sample surfaces and the lower part of lens；
- " Mag " is amplification factor；
- " inclination angle " represents the normal of sample stage relative to the angle of the axis of electron gun；And
- " signal A=SE2 " instructions use the detector of secondary electron.
With reference to figure 4：In embodiment, working lining 10 includes one or more semiconductor elements of such as silicon, and chamber 20
Pyramid shape there is the geometry consistent with the manufacturing process for obtaining chamber, use anisotropy the process advan
Etch process is (typically using having<100>The silicon chip of crystal orientation, the reason is that other orientations, such as<111>It will not produce desired
Pyramid shape).Now, it is considered below to be suitable for other semiconductors, such as Group IV element, example although silicon is preferable
Such as Ge or compound semiconductor, such as SiGe, other compound III-V or II-VI material, and their corresponding oxide
Or nitride.For example, GaAs and Ge can be etched anisotropically through.Moreover, this method the principle on which can be applied to one
A little metal layers and corresponding oxide.However, metal layer is less practical.Especially, they may not have phase in layer thickness
When crystal homogeneity.Furthermore, it is possible to for example expection uses Al2O3 surfaces.Al2O3 may be used as reaching the thin of 100-200nm
Film dielectric, and be deposited by sputtering or by atomic layer deposition ALD.The latter is expensive but high quality technology.Moreover,
Thickness for the layer needed for microballon trapping using ALD for having challenge.
Pyramid shape is unexpectedly compatible with anisotropic etching process.For Si or similar semiconductor element, chamber is manufactured
Anisotropic etching process cause 54.7 ° of desirable angle between the planar base portion (that is, be open 28) of pyramid and adjacent wall, this
Depth-width ratio of pyramid is determined again.It can be seen in fig. 4 that such technique allows the easy of chamber and cleaning manufacture.So
And the desirable angle may change slightly due to small defect.
In modification, dry film photoresist and more generally can be used, it is contemplated that some polymer, plastics and gold
Belong to.Preferably, using epoxy group dry film photoresist, it can be patterned by hot padding.Hot padding can also be applied to it
Its plastic material (for example, PMMA, COC, makrolon), may be not suitable for for the common plastics patterning method of the material
In generation pyramid microcavity.For example, after the array of anisotropic etching manufacture chamber of Si is used, can be electroplated by Ni
Carry out filled cavity.Electroplated Ni layer and then disengaging, form the mould or stamp that will be used in subsequent thermal imprint process.It can pass through
Pressure and temperature is applied to the stamp being in contact with plastics and then cools down and demoulds and Si chambers are produced on plastic layer
True copy thing.
Referring now more particularly to Figure 14, in embodiment, preparation chip causes its chamber (that is, at least one in chamber 20
A bit, preferably largely) it is filled with pearl 50,51,52.As it was earlier mentioned, such pearl is preferably microballon, i.e., have on average
There is the diameter between 1 to 40 μm.For application as contemplated herein, between 2 to 20 μm and more preferably between 2 to 10 μm
Diameter be preferable.Pearl is preferably polymeric beads, such as polystyrene bead, but can also use in principle silica beads or
Latex bead.
As it is shown in the figures, size of the size of the cross-sectional area of chamber equivalent to pearl.Pearl trapping chamber is to be designed specifically to
The trap of the single pearl of trapping.Therefore, pearl can be typically in two (or more) point contact cavity walls, or even be adhered to the point of chamber
At end." chamber " is contemplated to be the object different from microchannel or other microfluidic features herein.Due to solution presented herein
Certainly scheme, the most of of chamber 20 in array 30 can be filled with only one pearl 50,51,52, as seen in Figure 14.
At present, the details to the relative size of pearl on chamber is provided, it allows the stability of pearl and fall into that optimization is trapped
The occupancy of trap.Refer more especially to Fig. 3,11 and 15：Average (linear) size of chamber opening 28 and the ratio of average bead diameter
Should be preferably between 1.0 to 2.4.Moreover, have observed that optimum (in occupancy for the ratio between 1.4 to 2.4
Aspect) (it is, average 40-60% occupancies).When ratio is in 1.4-2.0, these results, which further improve, (is rinsing
Afterwards it is observed that averagely reaching 60% occupancy).In some cases, can obtain reach 90% occupancy.Moreover,
Above ratio is maintained at less than 2.0 or preferably less than the 1.8 multiple occupancy for allowing substantially to prevent same chamber.
For example, and the opening 28 of chamber 20 is limited as a result, for 10 μm of (diameter) pearls according to collect various of inventor
The full-size of planar base portion should be preferably less than 24 μm, and even more preferably less than 18 μm.These digit preferences for example by
Fluoroscopic image shown in Figure 14 confirms.The latter actually shows what the method for being trapped in and being described according to subsequent reference chart 5 obtained
The egative film of the fluoroscopic image of pearl in the array of chamber.Chip is obtained actually by the TMAH etchings for 13 μm of depths for performing Si substrates
Chamber；Use 200 μ m, 500 μm of arrays.By the way that (10 μm of Fluoro-MAX of about 200nL are come from Thermo
ScientificTM) pearl solution (undiluted) is pipetted on each array to integrate pearl.Therefore, Figure 14 image each
In, use the pearl of 10 μ m diameters.The lateral dimensions (invisible in the picture) of the opening of chamber is differed from 8 μm to 24 μm, and corresponding
Last occupancy rise to 63% from 5%, the peak value of 80%-90% is seen for 18-20 μm of perforate.For the specific feelings
Condition, the actual percentage of acquisition are：8μm：5.4%；10μm：24.4%；12μm：32.5%；14μm：46.3%；16μm：
51.6%；18μm：80.1%；20μm：89.9%；22μm：72.2%；And 24 μm：63.2%.However, multiple take occurs
Increase at 18 μm and herein.Therefore, therefore the scope of the size of perforate can be limited to (for 10 μ m diameter pearls)
Such as 14-18 μm, or even 16-18 μm.As one man, the average-size of chamber opening 28 and the ratio of average bead diameter can be limited
To 1.4-1.8 or 1.6-1.8.It should be noted that the percent value as caused by above particular case is different from previously mentioned average
Value, the reason is that the latter collects from various experiences.
Referring now to Fig. 3 and Figure 15：The ratio of the mean depth of chamber 20 and the average diameter of pearl 50 is preferably at least
0.5, more preferably 1.0, and be more preferably 1.3.For example, 10 μ m diameters are shown by the experiment that the present inventor carries out
Pearl can keep stablizing in 8 μ m, 8 μm of openings, but last occupancy/stability of discovery pearl is not good enough in this case
's.When using from the anisotropic etching process limited, such size means about 5 μm of depth.Therefore, average cavity depth
Ratio with average bead diameter can be with as low as 0.5.Now, for the cone through anisotropic etching, when it is desirable that obtaining bag completely
The depth of at least (about) 1.0d (wherein d is the diameter of pearl) is necessary when burying the pearl of (that is, embedded) in chamber, it is assumed that taper
Chamber is truncated and truncation surface 29 is sufficiently large to accommodate pearl, as shown in Figure 15.For example, 10 μm of pearls can be fully embedded to chamber
In, if opening is at least 19.33 μm of 19.33 μ m, and minimum at least 10 μm of depth, base etches during use；When base
Stop etching in etching in all planes before apex merging.As fruit caving is not truncated, (they have punctiform vertex, i.e., certainly
Limit etching), then ratio needs to be at least about 1.3, as shown in Figure 3.
Moreover, inventors have also recognized that pearl is without the need for being fully embedded in chamber.That is, a part for pearl can be with
Occur (that is, prominent) above the plane of opening 28 to keep satisfied stability, as shown in Figure 5 (not in scale).Therefore
Above-mentioned ratio can correspondingly be reduced.
Referring now to Fig. 1, Fig. 2, Fig. 6-9 and Figure 12-13, in embodiment, micro-fluid chip 100 can have chamber 20
Row or column in two or more subgroups 32, such as array 30, two of which subgroup are connected by least one microchannel 14.It is excellent
One or more of selection of land, these subgroups is defined in channel portion 12a, the bottom wall or roof of the channel portion by
Working lining 10 limits.Such configuration allows to multiplex, while prevents the cross contamination between subgroup 32.It is preferred, therefore, that
It is the subgroup 32 that array 30 is divided into pearl 50, to be partially isolating subgroup.The array of chamber can for example be arranged to match micro
Titrate the canonical coordinates in hole：On microtiter plate, hole is with the positioned at intervals of 9,4.5 or 2.25mm on secondary dot matrix.This can
In order to use the solution of robotic distribution pearl.In order to which the increase of the array of chamber is integrated, hexagonal-lattice can also be selected.
For example, used in the experiment shown in Figure 14 chip design include array, its along snakelike microfluidic channel (it is 200 μm wide, 14
μm depth) arranged and be distributed with hexagonal-lattice, wherein being about 1.1mm from an array to another angle diagonal distance.The arrangement
Allow to place 10 single arrays (each to take 200 μ m, 500 μm of areas) in the gross area of 3mm × 5mm, and allow
By the liquid droplet distribution of 200nL pearl solution to each array without merging drop or cross contamination.
Then, seen in " last " device as shown in figures 5-9, the array 30 of the chamber 20 of micro-fluid chip 100 is preferably
Sealed by the cap rock 60 extended relative to array 30.Cap rock is protected and annular seal space and their content.In addition to chamber 20,
Cap rock 60 will likely seal the other microfluidic structures being typically found on chip 100.The example of such micro-structure is：Load
Pad 11, detects antibody (or dAb) deposition region 70, capillary pump 16 or venthole 18, as shown in figs. 1 and 2.
It should be noted that although cap rock 60 extends relative to array 30, but it need not contact chamber, the reason is that it is generally necessary to
Gap is to form microfluidic channel.In the manufacturing process of Fig. 9, chamber is sealed by cap rock, the reason is that microchannel is arranged between chamber.And
And in other manufacturing technologies herein, epiphragma 60 is actually not directly contacted with chamber, always has and is deposited by channel layer
The gap that (Fig. 6) or etching (Fig. 7 and 8) introduce.Typically, channel depth is between 1 to 20 μm.More shallow passage will produce more
High flowed friction and the thickness evenness of smaller, and the manufacture of more deep channel will be more difficult and/or time-consuming.In general, can
With the minimum lateral size of the feature of manufacture since the aspect ratio limitation of passage manufacturing technology increases with the depth of passage.Mesh
Preceding manufacturing technology (wet etching or dry ecthing or dry film photoresist patterning) will be provided easily with the aspect ratio higher than 1
Structure (for example, 20 μm of wide features in the passage of 20 μm of depths) is without substantial amounts of parameter optimization effort.
Thus, channel height (or depth) can be adjusted to ensure that pearl will not depart from during flow.Due to this
Solution, the stability of pearl significantly improve.Moreover, may be always in the case where the flowing of high current body is present in microchannel
There is the residual risk for loosening some pearls.Now, if microfluidic channel depth is slightly less than the diameter of pearl (for example, example previous
It is less than 10 μm in son), pearl will never depart from during flow.However, too small channel depth (for example, 1 μm) will increase hydraulic pressure
Resistance and the total liquid capacity (volume) for reducing chip；Increase is loosened some pearls by too big depth (for example, more than 10 μm)
Probability.In the situation for removing excessive pearl using bringing, (that is, the pearl in the inside of chamber will retain and in the excess of channel interior
Band will be adhered to) under there is the channel depth of diameter for being slightly less than pearl also will be beneficial.
Referring now more particularly to Figure 12 and Figure 13, chip 100 can actually include some arrays 30 (each array bag
Include one or more pearl trapping chambers 20).These arrays 30 are preferably inserted in different pairs of microchannel in flow path is specified
Between part, i.e., in parallel or series.For example, these channel parts can be significantly made of parallel split tunnel, in Figure 13
It is shown, so that avoid from a part to another cross contamination, or can be arranged in series (such as in Figure 12).They
It can be arranged in the channel part of serpentine channel (not shown).
Then, it is possibility to have sharply using positioned at (one or more) array 30 chamber 20 in different types of pearl 51,
52.Due to preferred deposition technique, pearl 51,52 is typically deposited in different arrays 30.It is, for example, possible to use two kinds of different types
Pearl：A type of pearl 51 is used to analyze analyte detection, and another type of pearl 52 is used to compare, as shown in Figure 12,13.Such as
Previously described, depending on the practical application sought, array 30 can be with serial or parallel connection.It is expected most using (example herein
Such as, bioanalysis) in, pearl is different in terms of coating.But pearl can be more generally in size, coating, material and/or color side
Face is different.
Sealed it should be noted that the solution of the prior art is focused principally on before pearl (being flowed via pearl solution) is loaded
Device.This, which makes it difficult to (if possible), has the different types of pearl in the precalculated position being located in device.Therefore, this is right
More steps is actually meant that in bioanalysis.
According on the other hand, the present invention can be specifically embodied as the manufacture method of chip 100 as described above.Referring now to
Fig. 5-10 describes manufacture method.Most typically, such method is around the manufacture (step in the working lining 10,60 of device 100
S20) the basic step of the array 30 of pearl trapping chamber 20.With previously described device 100 as one man, this manufacture ensures in chamber 20
Each there is the taper that is limited by one or more side wall 21-24, the side wall is hydrophilic.Chamber 20 is used as blind hole
Extend in the thickness of working lining 10,60.
Depending on selected material 10,60, distinct methods are possible.Chamber can be located in substrate layer 10 or cap rock or
In film 60.As previously it is further described that manufacture method can include anisotropic etching process, thermal imprint process or any other
Suitable technique is to obtain chamber 20.So far, anisotropic etching process in terms of the quality obtained for chamber seemingly most
It is promising.Base etch process when being better than from the anisotropic etching process of limit, the reason is that the former is not easily susceptible to etch-rate
Change and allowable depth is kept nearly constant (last depth is limited by the size of chamber opening 28) in the case of overetched.
As it was earlier mentioned, anisotropic etching process preferably exists<100>Performed on chip,<100>Chip has<
110>Flat part on direction；Therefore top surface has<100>Normal on direction.The exposure of chip is accordingly parallel to
(100) plane, i.e., be orthogonal to (100) direction (being diamond structures for Si) on the basis of reciprocal lattice vector.Except chamber
Outside manufacture, anisotropic etching process can be used for manufacture microfluidic structures (for example, Fig. 8 and 9).If there is inclined side
Wall is not harmful for microfluidic structures (such as passage), then has<100>The wet etching of the Si chips of crystal orientation is better than dry corrosion
Lithography, the reason is that wet etching is compatible with batch processing and therefore can generally faster, this depends on processed chip
Quantity.It should be noted that wet etching is usually slower than dry ecthing, each chip is produced, dry ecthing is faster.Therefore total output takes
Certainly in the quantity for the chip processed together.
If necessary, (one or more) array 30 can be cleaned (for example, using ethanol, water etc.) and/or use plasma
Body (for example, air, oxygen or helium) processing.In all cases it is possible to only by by the droplet spreads S32 of pearl solution 55 to example
Pearl 50,51,52 is such as deposited into S30 into chamber 20 on the top of (one or more) array 30.For example, can will about 2 μ L
Deposit pearl solution (being provided typically as 1% solid suspension) be applied to (one or more) array 30.
(and before S40 chips are sealed) after depositing S30 pearls, it is proposed that：
- dry S34 chambers 20 (pearl 50 be maintained in chamber or near)；And then
- for example rinse S36a (one or more) array 30 by using rinse solution and/or be used to adhere to by applying
Bringing for S36b excess pearls removes S36 excess pearl (that is, not being trapped in those pearls inside chamber 20)；Afterwards
- if necessary, dry S38 again has the array 30 of chamber 20 of remaining pearl wherein, i.e., does not always need finally dry
Dry step, such as when only applying band to remove excessive pearl.
Present inventors have recognized that dried array 30 unexpectedly causes more stable pearl before excessive pearl is removed
50.Consequently found that the influence to last occupancy is significant：Depending on other conditions, due to first drying steps S34, account for
It can improve 20% to 60% with rate, it is even more.It may infer that conical cavity is more preferable when flushing and dry (for example, N2 flows)
Protect pearl from lateral, move in plane the influence of fluid (gas or liquid) in ground.Preferable rinse solution is, for example, to buffer
Solution or deionized water.More generally, it can be any solution that can not adversely influence pearl or the albumen being coated on.
Finally, array 30 (more generally part or all of chip 100) can use cap rock 60 (for example, dry film) to seal
S40, the cap rock are preferably laminated to ensure excellent sealing.When chamber is located in layer 10, lid 60 is positioned to relative to chamber 20
Array 30 extend, be consequently formed closure microfluidic channel 12,12a, 14 and structure 16,18.
Discuss four different manufacture examples in detail now.First manufacture example is illustrated in detail in Figure 5.Fig. 5's is detailed
In addition thin step is captured in the flow chart of Figure 10.Fig. 6 only shows the selected step of first manufacture method, and Fig. 7-9
The selected step of other possible manufacture methods is shown.In each of Fig. 6-9, upper row's figure is the miniflow in the different fabrication stages
The sectional view of body chip, and lower row shows corresponding top view.
With reference first to Fig. 5, Fig. 6 and Figure 10, the first manufacture example is etched using anisotropy chamber, then against corrosion using dry film
Agent passage manufactures.In detail：
- step S10：Micro-fluid chip main body is provided, it includes layer (or substrate) 10.
- block S20：The manufacture of chamber and passage：
Zero S21, Fig. 5 A：Layer 10 is aoxidized, for example, silicon is aoxidized by thermal oxide to obtain SiO2 layers of 10o.The electricity of acquisition
Insulating layer 10o typically covers whole substrate 10.Instead of oxide, can also attempt to obtain nitride, such as Si3N4.
Zero in step S22, Fig. 5 B：Such as use dry and wet etching or wet etching patterned oxide.For this purpose, it is photic
Resist typically serves as mask (Fig. 5 B are schematically depicted in etching oxide and peel off the chip after photoresist)；
Zero then, anisotropic etching substrate layer 10, step S24, Fig. 5 C, so as to obtain chamber 20 and other deposition regions
70.Preferably, using wet etchant, it is typically TMAH or KOH.Typically short oxide etching is carried out before Si etchings
(such as BHF) is to remove the native oxide on Si surfaces；
Zero then：Peel off oxide, step S25, Fig. 5 D.But this is optional, the reason is that oxide can be with when needing
Remain on the surface.Typically, buffer oxide etch (such as BHF) is for the purpose；And preferably by depositing, exposure and
Then develop dry film photoresist (negative photoresist) or epoxy-based negative photoresist (such as SU-8) patterning passage
Side wall 62, step S26, Fig. 5 D；
- block S30：Pearl deposition (also referred to as reagent is integrated)：
Zero step S32, Fig. 5 E：(different types of pearl can be deposited on phase to the droplet deposition of pearl solution on chamber array 20
On the array answered, as shown in Figures 12-13)；
Zero step S34, Fig. 5 F：Dried array 30 and pearl (preferably by natural evaporation, by the air-flow of N2 or pass through by
Chip is placed in controlled environment or in warm plate, etc.)；
Zero step S36：Excessive pearl is removed, is preferably passed through：(i) them are rinsed, for example, it is molten using deionized water or buffering
The stream of liquid, step S36a, Fig. 5 G；And/or (ii) applies the band of such as adhesive tape or PDMS, excessive pearl will adhere to the band, walk
Rapid S36b, Fig. 5 H.If necessary, band is repeatedly applied；And
- S40, Fig. 5 I：Finally, chip 100 (one or more) array 30 of chamber (particularly) is with being preferably laminated S42
Cap rock 60 (for example, dry film photoresist) sealing.For this purpose, cap rock 60 typically by appropriate heating to such as 45-50 DEG C.
Epiphragma can have (to be seen, Fig. 6) in step s 40 in the opening loaded at pad with liquid relief liquid.Can be by cutting or rushing
Press to pattern such opening.
The example above of manufacture method allows flexible design, the reason is that the manufacture of chamber and passage is separated.It can also recognize
Arrive, such method allows circular passageway configuration.But the defects of such method, is that its needs mask and is rushed twice
The pearl washed may stay in channel interior.
Referring now to Figure 7, the manufacture example of description second, wherein integument be incorporated into cap rock 60 (for example, dry film photoresist) and
It is not layer 10.Briefly：
- step S20a：Using mould 65 with the patterning chamber (for example, passing through hot padding) in layer 60；
- step S20b：Separate patterned layer 60；
- step S30：Pearl is deposited on again by liquid relief pearl solution droplets, drying, flushing and drying again patterned
In the chamber of layer 60；
- step S40：Finally, it is preferably close thereon by the way that cap rock 60 is laminated to by the way that cap rock 60 is placed on chip
Seal MF chips.
In the method, the integration of pearl is unrelated with substrate and passage, this allows the bigger spirit of the design of substrate 10 and passage
Activity.MF chips, which can have, is etched (anisotropic etching, or deep reaction etching) or deposition (dry film photoresist, SU-8
Deng) or pass through hot padding or the microfluidic structures of moulded pattern.Epiphragma, which can be laminated to, is with or without advance surface modification
Or on any compliant substrate of processing.However, this method may need transparent substrates for more effective fluoroscopic examination.And
And compared to the pearl being incorporated into substrate, pearl may be easier to depart from from film.
Then, with reference to figure 8, the 3rd manufacture example is easily described, it is etched using two step photoetching and anisotropy Si：
- step S20c：Etched by anisotropy Si come etched channels.Then thermal oxide (not shown) is carried out；Then
- step S20d：Etched after the patterning of oxide by anisotropy Si come etched cavity；
- step S30：Integrate pearl；And
- step S40：Encapsulating chip.
Such manufacture method allows flexible design, the reason is that trap and passage manufacture are separated.However, it is needed more
Manufacturing step (needing mask twice), and the additional thermal oxide between two etching steps.In addition, in the method, quilt
The pearl of flushing may stay in channel interior.
Finally, and with reference to figure 9, last manufacture example is described, it is etched dependent on single step photolithography and anisotropy Si.
Step S20e：Anisotropic etching etched channels and chamber in a single photolithographic step are used in same time.Then, in step
S30 integrates pearl, and encapsulating chip, step S40.Using such method, passage and trap are manufactured in an etching step
Both, its cost benefit higher.Moreover, passage depth can be limited by adjusting the width of passage in the etch process from limit
Degree, the reason is that broader opening causes deeper passage, vice versa.Each chamber can be connected to each other simultaneously via microfluidic channel
And microfluidic networks are connected to, the width of the microfluidic channel is less than the width of chamber, as shown in S20e (Fig. 9).However, this
The manufacture method of sample means tightened up design rule, since the small size for the microfluidic channel for interconnecting chamber causes higher
Flowed friction.Also there is the risk for being internally formed bubble in chamber.
At present, the example of the application of the embodiment of the present invention is discussed.It is, in open channel system (in order to simple
For the sake of, the sealing of chip unused epiphragma) in the simple ligand-receptor measure of demonstration：
Using from Bangs Laboratories Inc.TMPolystyrene " Superavidin " coating pearl
(10 μ m diameter)：10 μm of polystyrene are coated with avidin (a kind of known albumen for being attached to very much biotin by force)
Pearl.Commercial stock solution includes the pearl as 1% solid；
Stock solution is diluted to 1/5 (dilution that can also use bigger) with PBS+0.5% polysorbas20s；
Use the array of the chamber obtained according to Fig. 5 A-5C (the step S10 to S24 in Figure 10)：The pearl of about 2 μ L is molten
Liquid is dispersed on array and is dried and (is enough within 1 to 2 minutes to complete drying)；
Array and pearl are rinsed in flowing down for PBS+0.5% polysorbas20s (about 30mL, continue 10s)；
Then array and pearl are rinsed under DI current (about 30mL, continue 10s)；
Then the dried array under nitrogen stream；
Array is covered with the drop of the 1%BSA+0.5% polysorbas20s in PBS and pearl continues 15 minutes；
Array and pearl and the drying under nitrogen stream are rinsed with PBS+0.5% polysorbas20s, water.
Finally, can seal and storage chip.
Measured for typical case, user carries out following three steps, and wherein analyte is represented by fluorescence-labeled bio element molecule.
Array and pearl are exposed to 50 μ g/mL biotins -590-AttoSolution continue
15 minutes, while it is protected from light.During the step, biotin -590-Atto is attached to the avidin on pearl；
Array and pearl are rinsed with PBS+0.5% polysorbas20s and water.These rinsing steps are optional, but can be answered
With to increase the sensitivity of measure.
Using fluorescence microscope monitoring biotin-Atto-590 (ligand) be attached on pearl avidin (by
Body).Single pearl chamber allows simple signal-obtaining and explains without any optical artifacts.
Method and apparatus specifically described herein can use in the manufacture of micro-fluid chip.Resulting chip can be with
By manufacturer in original form (for example, bilayer device as structuring) or with packaged form distribution.In the latter case
Chip may be mounted in single-chip package.Under any circumstance then chip can be integrated with other elements, as among (a)
A part for product or (b) final products.
In short, the embodiment of the present invention provides various advantages.Such as：
- the embodiment of the present invention allows to keep pearl in a manner of especially and unexpectedly stablizing, if especially exist
Dry beads and chamber array before the excessive pearl of removal.A small amount of pearl is only lost during liquid and drying is subsequently exposed to.To with
After the research of the whole world for many years of the microfluidic device and patterning acceptor of measure, find to be used to integrate pearl (or acceptor) finally
To the very effective ways of microfluidic device；
- current design and corresponding manufacture method also allow to place pearl before trap is covered；
The array of-chamber can determine size so as to each chamber trapping list pearl relative to pearl.Single pearl chamber allows simpler letter
Number read and explain, it is necessary to which individual layer is to accommodate pearl, and causes less (even without) optical artifacts；And
- manufacture method presented herein allows high production rate to manufacture.Various pearl integrated strategies are suggested, it allows miniflow
The flexible design of body device and flow path.The concept that the embodiment of the present invention is based on by accident with some microfluidic features
(mirror, plastic chip manufacture etc.) is compatible.
Although with reference to the embodiment of limited quantity, modification and the attached drawing description present invention, those skilled in the art
It will be understood that various change can be carried out and equivalent can be replaced without departing from the scope of the present invention.Especially, specifying
Described in embodiment, modification or the feature (device or method) that is shown in figure can with another embodiment, modification or figure
Another feature combines or replaces another feature, without departing from the scope of the present invention.Therefore it is anticipated that on above example or
The various combinations of feature described in any one in modification, it is still in the range of attached claims.Furthermore it is possible to
Many small modifications are carried out so that particular case or material are adapted to scope of the teachings of the present invention without departing from it.So
It is contemplated that disclosed specific embodiment is unlimited to, but the present invention is by including falling the scope in attached claims
Interior all embodiments.Furthermore it is possible to expect to be different from many other modifications being explicitly described above.For example, different from herein
In the other materials that are expressly recited can be used for layer 10,60 each.In addition, passage, load pad, venthole, chamber etc. can be with
With various sizes.
Reference numerals list
10 basic units
10o oxide skin(coating)s
11 load pad
12 microfluidic channels (microchannel)
12a channel portions
14 multiplexing microchannels
16 capillary pumps
18 ventholes
20 pearl trapping chambers
The side wall of 21-24 chambers
The wall of 24a-d structurings (multiaspect)
28 chamber openings/tapered base
29 bottom of chamber surface
The array of 30 pearl trapping chambers
32 thorax groups (row/column)
50 pearls
51 analysis analyte detection pearls
52 control pearls
55 pearl solution (drop)
60 cap rocks (dry film), lid
62 conduit walls
65 are used for the mould of the coining chamber in cap rock 60
70 are used to detect the chamber that antibody (dAb) deposits
The detection antibody (dAb) of 72 depositions
100 micro-fluid chips
Claims (23)
1. a kind of micro-fluid chip (100), including layer (10,60), the array (30) of pearl trapping chamber (20) be located at the layer (10,
60) in, wherein each in the chamber (20) has the taper limited by one or more side walls, it is characterised in that described
Side wall is hydrophilic, and each in wherein described chamber (20) is prolonged as blind hole in the thickness of the layer (10,60)
Stretch.
2. micro-fluid chip (100) according to claim 1, wherein at least some with by side wall in the chamber (20)
The pyramid shape of formation, the side wall are hydrophilic, and wherein, which is substantially limited by least four sidewalls
It is fixed.
3. micro-fluid chip (100) according to claim 1 or 2, partly leads wherein the layer (10) includes one or more
Volume elements part, and the pyramid shape of the chamber (20) has the anisotropic etching with the manufacture of the chamber in the layer (10)
The consistent geometry of technique.
4. micro-fluid chip (100) according to claim 1, wherein at least one in the chamber (20) is filled with pearl,
The pearl averagely has the diameter between 1 to 40 μm.
5. micro-fluid chip (100) according to claim 4, wherein the pearl averagely has the diameter between 2 to 20 μm.
6. micro-fluid chip (100) according to claim 4, wherein the pearl averagely has the diameter between 2 to 10 μm.
7. the micro-fluid chip (100) according to any one of claim 4-6, wherein the chamber (20) of the array (30)
In major part be filled with only one pearl.
8. the micro-fluid chip (100) according to any one of claim 4-6, wherein the opening (28) of the chamber (20)
Average-size and the pearl in the chamber (20) average diameter ratio between 1.0 to 2.4.
9. micro-fluid chip (100) according to claim 8, wherein the average-size of the opening (28) of the chamber (20) with
The ratio of the average diameter of pearl in the chamber (20) is between 1.4 to 2.0.
10. the micro-fluid chip (100) according to any one of claim 4 to 6, wherein the average depth of the chamber (20)
Degree and the ratio of the average diameter of the pearl in the chamber (20) are at least 0.5.
11. micro-fluid chip (100) according to claim 10, wherein the mean depth of the chamber (20) and the chamber
(20) ratio of the average diameter of the pearl in is at least 1.0.
12. micro-fluid chip (100) according to claim 10, wherein the mean depth of the chamber (20) and the chamber
(20) ratio of the average diameter of the pearl in is at least 1.3.
13. micro-fluid chip (100) according to claim 1, two of which or more is by least one chamber (20) structure
Into subgroup (32) by least one microchannel (14) connect.
14. micro-fluid chip (100) according to claim 13, wherein the subgroup is the array (30) of the chamber (20)
In row or column.
15. the micro-fluid chip (100) according to claim 13 or 14, wherein one or more of described subgroup is micro-
It is defined in channel part (12a), the bottom wall or roof of the channel portion are formed by the surface of the layer (10).
16. micro-fluid chip (100) according to claim 1, wherein the array (30) is by relative to the chamber (20)
Array (30) extension cap rock (60) sealing.
17. micro-fluid chip (100) according to claim 1, includes some battle arrays of one or more pearl trapping chambers (20)
Arrange (30), wherein these arrays are inserted between paired channel portion (14).
18. micro-fluid chip (100) according to claim 17, including be correspondingly situated in some arrays (30)
At least two different types of pearls in one or more chambers of at least two arrays.
19. micro-fluid chip (100) according to claim 18, wherein at least two different types of pearl is in ruler
It is different in terms of very little, coating, material and/or color.
20. a kind of manufacture method of micro-fluid chip (100) according to any one of claim 1 to 19, including：
There is provided (S10) has the micro-fluid chip main body of layer (10,60)；And
The array (30) of manufacture (S20) pearl trapping chamber (20) in the layer (10,60), wherein each tool of the chamber (20)
There is the taper limited by one or more side walls, the side wall is hydrophilic, and each work of wherein described chamber (20)
Extend for blind hole in the thickness of the layer (10,60).
21. according to the method for claim 20, wherein manufacture (S20) described array (30) is included the use of from each to different of limit
Property etch process is etched anisotropically through (S24) described layer (10) to obtain the chamber (20).
22. the method according to claim 20 or 21, further includes：
Pearl is deposited into the chamber (20) of the array (30) by (S30) by the drop for spreading (S32) pearl solution (55)；With
And
Cap rock (60) sealing (S40) the described array (30) extended with the array (30) being positioned to relative to the chamber (20), and
And (S40) described array (30) is wherein sealed including being laminated (S42) described cap rock (60).
It is 23. described according to the method for claim 22, wherein after deposition (S30) pearl and before (S40) is sealed
Method further includes in the following order：
Dry (S34) has the array (30) of the chamber (20) of pearl wherein；
(S36a) described array (30) is rinsed by using rinse solution and/or by applying the band for being used for adhering to (S36b) excessive pearl
Remove the excessive pearl that (S36) is not trapped in the inside of the chamber (20)；And
Dry (S38) has the array (30) of the chamber (20) of pearl wherein again.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB1405402.7A GB2524541A (en) | 2014-03-26 | 2014-03-26 | Microfluidic chip with conic bead trapping cavities and fabrication thereof |
| GB1405402.7 | 2014-03-26 | ||
| PCT/IB2015/051579 WO2015145280A1 (en) | 2014-03-26 | 2015-03-04 | Microfluidic chip with conic bead trapping cavities and fabrication thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106104271A CN106104271A (en) | 2016-11-09 |
| CN106104271B true CN106104271B (en) | 2018-04-20 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201580012689.2A Expired - Fee Related CN106104271B (en) | 2014-03-26 | 2015-03-04 | Micro-fluid chip and its manufacture with conical bead trapping chamber |
Country Status (5)
| Country | Link |
|---|---|
| JP (1) | JP2017512987A (en) |
| CN (1) | CN106104271B (en) |
| DE (1) | DE112015000920B4 (en) |
| GB (2) | GB2524541A (en) |
| WO (1) | WO2015145280A1 (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB201610426D0 (en) | 2016-06-15 | 2016-07-27 | Q-Linea Ab | Sample holder for image based analysis of samples |
| WO2018175500A1 (en) * | 2017-03-21 | 2018-09-27 | Hexanomics, Inc. | Sealed microwell assay |
| CN107583692B (en) * | 2017-05-23 | 2022-11-11 | 深圳市博瑞生物科技有限公司 | Liquid drop micro-fluidic chip and preparation method thereof |
| EP3704058A4 (en) * | 2017-10-31 | 2021-11-10 | Astrego Diagnostics AB | Microfluidic device for cell characterization |
| DE102018203047A1 (en) * | 2018-03-01 | 2019-09-05 | Robert Bosch Gmbh | Microfluidic device |
| WO2020062149A1 (en) * | 2018-09-29 | 2020-04-02 | 天津大学 | Flexible encapsulation structure and manufacturing method therefor |
| CN109701671A (en) * | 2018-12-27 | 2019-05-03 | 复旦大学 | A kind of microlayer model array chip and production and preparation method thereof |
| US11173486B2 (en) | 2019-02-13 | 2021-11-16 | International Business Machines Corporation | Fluidic cavities for on-chip layering and sealing of separation arrays |
| CN112723303B (en) * | 2020-12-14 | 2021-09-24 | 苏州拉索生物芯片科技有限公司 | Microbead chip and spin coating preparation method thereof |
| US20230132688A1 (en) * | 2021-10-28 | 2023-05-04 | Worcester Polytechnic Institute | Gravity independent liquid cooling for electronics |
| CN114308163B (en) * | 2021-12-31 | 2024-01-09 | 北京京东方技术开发有限公司 | Microfluidic chip detection cartridge |
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- 2015-03-04 CN CN201580012689.2A patent/CN106104271B/en not_active Expired - Fee Related
- 2015-03-04 GB GB1617833.7A patent/GB2548941B/en active Active
- 2015-03-04 JP JP2016556828A patent/JP2017512987A/en active Pending
- 2015-03-04 WO PCT/IB2015/051579 patent/WO2015145280A1/en active Application Filing
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| JPH10332593A (en) * | 1997-05-27 | 1998-12-18 | Hitachi Ltd | Electrochemiluminescence detecting cell |
| US6908770B1 (en) * | 1998-07-16 | 2005-06-21 | Board Of Regents, The University Of Texas System | Fluid based analysis of multiple analytes by a sensor array |
| CN1267089A (en) * | 1999-03-15 | 2000-09-20 | 清华大学 | Single-point strobed micro electromagnetic units array chip or electromagnetic biologic chip and application thereof |
| US6589779B1 (en) * | 1999-07-16 | 2003-07-08 | Board Of Regents, The University Of Texas System | General signaling protocol for chemical receptors in immobilized matrices |
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Also Published As
| Publication number | Publication date |
|---|---|
| GB201617833D0 (en) | 2016-12-07 |
| JP2017512987A (en) | 2017-05-25 |
| CN106104271A (en) | 2016-11-09 |
| DE112015000920T5 (en) | 2016-11-24 |
| GB2548941A (en) | 2017-10-04 |
| GB2524541A (en) | 2015-09-30 |
| GB2548941B (en) | 2020-08-19 |
| DE112015000920B4 (en) | 2021-09-30 |
| GB201405402D0 (en) | 2014-05-07 |
| WO2015145280A1 (en) | 2015-10-01 |
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