CN106256436B - The micro flow control chip device and method of the anti-drop evaporation of channel interval formula - Google Patents
The micro flow control chip device and method of the anti-drop evaporation of channel interval formula Download PDFInfo
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- CN106256436B CN106256436B CN201610613672.9A CN201610613672A CN106256436B CN 106256436 B CN106256436 B CN 106256436B CN 201610613672 A CN201610613672 A CN 201610613672A CN 106256436 B CN106256436 B CN 106256436B
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- drop
- vaporization prevention
- microlayer model
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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
- B01L3/502769—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 multiphase flow arrangements
- B01L3/502784—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 multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
- B01L3/502792—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 multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics for moving individual droplets on a plate, e.g. by locally altering surface tension
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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
- B01L2200/0642—Filling fluids into wells by specific techniques
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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
- B01L2200/14—Process control and prevention of errors
- B01L2200/142—Preventing evaporation
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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
- B01L2300/06—Auxiliary integrated devices, integrated components
- B01L2300/0672—Integrated piercing tool
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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
- B01L2300/0809—Geometry, shape and general structure rectangular shaped
- B01L2300/0829—Multi-well plates; Microtitration plates
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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
- B01L2400/00—Moving or stopping fluids
- B01L2400/02—Drop detachment mechanisms of single droplets from nozzles or pins
Abstract
The invention discloses a kind of micro flow control chip devices of the anti-drop evaporation of channel interval formula, including：Manipulate probe；Drop plate for carrying microlayer model；Vaporization prevention cover board of the closed the cover on drop plate drop loading end, the position of drop zone described in the vaporization prevention cover board face are equipped with the through-hole structure when generating drop for avoiding manipulation probe；Confined space is formed between the vaporization prevention cover board and drop plate, and gas is filled in confined space；And after ensureing that microlayer model generates, there are external series gaps between the vaporization prevention phase and microlayer model；In the through-hole structure or through-hole structure top surface is sealed with vaporization prevention phase.The present invention is mutually not directly contacted with microlayer model using vaporization prevention, in the local confined space of formation using gas by between the two every method, the cross-contamination issue between microlayer model and vaporization prevention phase, microlayer model and microlayer model can be avoided completely, meanwhile and the vaporization prevention and interval problem of microlayer model can be solved.
Description
Technical field
Field of the present invention is microfluidic analysis field, is specifically related to a kind of anti-drop evaporation of channel interval formula
Micro flow control chip device and method.
Background technology
Drop microflow control technique is that a kind of tool occurred in micro-fluidic field in recent years grows a lot the new technologies of potentiality
And research direction.It simply says, micro-fluidic drop is a kind of technology that fine droplet is generated and manipulated in microchannel network.Area
Not in homogeneous continuous flow system, drop refer to a phase liquid be isolated with its immiscible another phase or multiphase to be formed it is non-
Continuous fluid.Compared with continuous flow microfluidic system, in liquid drop microfluidic system, it can be made by the immiscible drop being isolated
For individual microreactor, no dilution and cross contamination, drop internal sample mixed and heat in long-time is reacted and is transmitted
Transmission greatly accelerates that the reaction time is obviously reduced.The micro-fluidic important development direction as current microflow control technique of drop,
Its application field includes nano-particle, nano molecular self assembly, crystallization of protein, enzyme, DNA analysis, Cellular and subcellular structure
Analysis, high-flux medicaments sifting etc..
Currently, in liquid drop microfluidic system, be all using oil phase as between drop interval phase or package mutually to prevent
Between the evaporation of microlayer model, and different drop cross contamination (Brouzes E, Medkova M, Savenelli N, PNAS,
2009,106:14195-14200；Mazutis L,Gilbert J,Ung W L,Nat.Protoc.,2013,8:870-891；
Joensson H N,Svahn H A,Angew.Chem.Int.Edit,2012,51:12176-12192).But oil phase and liquid
Drop is in direct contact, and can also there are problems that：Drop internal part liposoluble substance is possible to be dissolved in the oil of interval aqueous phase droplets
Xiang Zhong thus will appear the cross-contamination issue between different drops.Therefore, how most under the premise of preventing microlayer model from evaporating
Cross contamination between amount avoids droplets from is current urgent problem to be solved.
Invention content
The object of the present invention is to provide a kind of using the micro drop of channel interval and vaporization prevention phase, and then prevents micro liquid
Drop evaporation while the micro flow control chip device for reducing cross contamination between drop.
The device of the invention combines the advantage of micro-fluidic chip and drop technique, using gas by drop and vaporization prevention phase
Separation had not only avoided Conventional drop and material exchange when vaporization prevention is in contact between drop, but also drop can be prevented because of evaporation
And volume reduces, and entire drop of testing liquid operation can be rapidly completed, and help to realize in-situ treatment and detection.Suitable for answering
For biochemical analysis, high-flux medicaments sifting, the protein crystallization condition screening of micro-scale volume liquid, cell analysis, even
Single cell analysis, single molecule analysis etc. need to carry out the occasion of micro liquid manipulation and reaction.
A kind of micro flow control chip device of the anti-drop evaporation of channel interval formula, including：
Manipulation probe for generating and manipulating microlayer model；
Drop plate for carrying microlayer model, the drop loading end of the drop plate, which is equipped with, is used to form one of microlayer model
Or multiple drop zones；One drop corresponds to a drop zone or a drop zone corresponds to several drops, can be according to practical reality
Test it needs to be determined that；
Vaporization prevention cover board of the closed the cover on drop plate drop loading end, drop zone described in the vaporization prevention cover board face
Position is equipped with the through-hole structure for being passed through for manipulating probe when generating or manipulating drop；
Confined space is formed between the vaporization prevention cover board and drop plate, and gas is filled in confined space；And ensure micro-
After drop formation, there are external series gaps between the vaporization prevention phase and microlayer model；
In the through-hole structure or through-hole structure top surface is sealed with vaporization prevention phase.
When drop zone is multiple, multiple microlayer models can share a confined space, preferably, the vaporization prevention lid
Plate realizes that sealing is fixed with drop plate by spacing board.In this scheme, spacing board, which plays, separates vaporization prevention cover board and drop plate
Effect, the confined space of an entirety is collectively formed with vaporization prevention cover board and drop plate.
Alternatively, each drop zone also can be mutual indepedent, there is the sealing space of oneself, as another preferred embodiment, single liquid
It drips area periphery and is equipped with spacing board, be tightly connected respectively with vaporization prevention cover board and drop plate at the top and bottom of spacing board, it is two neighboring
Drop zone is mutually isolated by the spacing board.In this technical solution, spacing board plays interval action, to the drop of its inside into
Row sealing provides confined space to each drop zone.
In the present invention, the confined space contains the gas for playing the role of being spaced vaporization prevention phase and microlayer model.This is closed
Space is the shared overall space of multiple microlayer models, either each microlayer model dedicated individual space or above two class alone
The various combination of the gas compartment.The volume range of confined space is 1 picoliters to 100 liters.Alternatively, preferably, the confined air
The interior protective gas for being filled with specific composition.
In the present invention, the mutual alignment relation of three-decker is vaporization prevention cover board, spacing board, drop plate；Three-decker it
Between combine closely, and ensure that gas cannot be leaked from the engaging portion of different layers structure；Microlayer model is located at by vaporization prevention cover board
It is full of in the confined space that (including vaporization prevention phase), spacing board, drop plate surround, in the confined space and plays interval vaporization prevention phase
With the gas of microlayer model effect；Manipulating probe can pass through vaporization prevention mutually to enter in confined space, to micro- liquid in confined space
Drop is manipulated.
Preferably, the drop zone is groove structure.When using the technical solution, spacing board can be saved, directly will
Vaporization prevention cover board and the sealing of drop plate are fixed, ensure that microlayer model and vaporization prevention are mutually mutual by the depth of adjusting grooves structure
Separation.
In the present invention, microlayer model is located at the surface of drop plate, either in the groove of drop plate surface or positioned at liquid
In the region limited by different hydrophobe properties surface on drop plate.Microlayer model can be accurately positioned and seek in the position of drop layer
Location.
A kind of Microfluidic droplet device using channel interval microlayer model and vaporization prevention phase of the present invention, micro- liquid
Drop can be the liquid of any form, type, mainly determine according to actual needs.These liquid include sample liquid or reaction solution,
Or buffer solution or the one or more or aqueous phase liquid in dilution or other functional solutions or oil phase liquid.Microlayer model
Volume range be that 1 Ah rises to 1 milliliter.
The vaporization prevention is mutually the liquid that can be avoided or reduce the gas evaporation from microlayer model of any form, type
Body.Specific type determines according to actual needs.Vaporization prevention is mutually housed inside in vaporization prevention layer, is not in direct contact with microlayer model.It is anti-
The composition of evaporation phase can be the oil phase containing organic principle or inorganic constituents, such as mineral oil, dimethicone, fluorocarbon oil, or
It is water phase, such as pure water or aqueous solution.Preferably, generally selecting uniform, stable, transparent, non-blooming light color liquid conduct
Vaporization prevention phase be judged by accident when in this way can be to avoid direct observation, will not occur being obstructed using road of micro- sem observation time or
Influence observed result.
According to the present invention, the vaporization prevention cover board, spacing board, drop plate material be can effectively prevent gas transmission consolidate
Body material can be glass, quartz, silicon, metal, high molecular polymer, inorganic or organic material and repressed fibrous material
Deng.It is equipped with the space (i.e. through-hole structure) of carrying vaporization prevention phase in the vaporization prevention cover board, is equipped in confined space and accommodates gas
The space of body.The thickness of above-mentioned vaporization prevention cover board and spacing board can be as thin as 1 micron, and thickness is to 20 centimetres.
The shape that vaporization prevention plate carries the horizontal cross-section in the space of vaporization prevention phase is circle, oval, rectangle or other arbitrary
Polygonal shape.The horizontal cross-section minimum at length range can be greatly to 1 centimetre, as low as 10 microns, can allow for probe into
Go out vaporization prevention phase, it is unobstructed, and the seal of vaporization prevention phase is not destroyed.Preventing preferably, the through-hole structure is setting
The mutually independent single hole arrangements of cover board are evaporated, each single hole arrangements are with the drop zone up and down to just.In the technical program, institute
The shape in the space of carrying vaporization prevention phase is through-hole shape structure in the vaporization prevention cover board stated, and is mutually held vaporization prevention using surface tension
It is loaded in through-hole.Multiple through-holes are processed on vaporization prevention cover board, to carry out the manipulation of multiple microlayer models.As another preferred embodiment,
The through-hole structure that vaporization prevention phase is carried on the vaporization prevention cover board is networked pore structures or comb shape pore structure, utilizes surface tension
Vaporization prevention is mutually carried in mesh or at the gap of comb shaped structure.
According to the present invention, the thickness of the drop plate can be as thin as 1 micron, and thickness is to 20 centimetres.Drop plate surface can be with
Cross sectional shape for the plane (groove structure) with pitting, hole can be round, oval, rectangle or any other polygon shape
Shape.The close and distant water process of selectivity can also be carried out in drop layer surface.The use of pitting and the close and distant method for treating water of selectivity be for
It controls and the position of fixed microlayer model, to avoid contact between different microlayer models and between microlayer model and vaporization prevention phase, production
Raw cross contamination.The microlayer model is located at the surface of drop plate, either in the pitting of drop plate surface or positioned at liquid
In the region limited by different hydrophobe properties surface on drop plate.Microlayer model can be accurately positioned and seek in the position of drop plate
Location.The volume range of microlayer model is that 1 Ah rises to 1000 microlitres.
Preferably, the manipulation probe for generating and manipulating microlayer model is the capillary-like structures with channel, or
There are probe-like structures that are specific function or being fixed with functional material for solid acicular texture or surface；In one system
The number for manipulating probe can more than one；The material for manipulating probe is inorganic material or organic material or high molecular polymer material
Material or organic-inorganic composite material.
Certainly, the property according to vaporization prevention lid surface property difference or vaporization prevention phase is different, and vaporization prevention is mutually
It can support in vaporization prevention cover board top surface, i.e., vaporization prevention phase the cover is in through-hole structure top surface.According to the present invention, by above-mentioned anti-steaming
It sends out the selection of the material of layer or selective hydrophilic and hydrophobic processing is carried out to its surface, more reliably can carry or fix anti-steaming
Send out phase.
According to the present invention, the function of the spacing board is to provide the gas for playing interval microlayer model and vaporization prevention phase function
Accommodation space.The confined space surrounded by vaporization prevention cover board (including vaporization prevention phase), spacing board, drop plate.The confined space
It is interior to be full of the gas for playing the role of being spaced vaporization prevention phase and microlayer model.The vaporization prevention cover board, spacing board, drop plate mutually it
Between junction must fit closely, or be sealed using adhesive etc., leaked to avoid gas.
Spacing board can not be used, directly by vaporization prevention when drop plate carries microlayer model with groove structure according to the present invention
Cover board (including vaporization prevention phase) and the hardened conjunction of drop, surround confined space.
The present invention also provides a kind of miniflows using the anti-drop evaporation of channel interval formula described in any of the above-described technical solution
The method that control chip apparatus carries out micro-fluidic operation, including：Using the liquid for manipulating probe carrying composition microlayer model, anti-steaming is passed through
Hair mutually enters in confined space, in the drop zone of drop plate surface, forms microlayer model.Or pass through anti-steaming using probe is manipulated
Hair mutually enters in confined space, liquid is added into the microlayer model formed, or by taking out liquid in microlayer model.
As a preferred embodiment, the method for carrying out micro-fluidic operation includes：
(a) vaporization prevention cover board (including vaporization prevention phase), spacing board, drop plate is utilized to surround confined space, in confined space
Full of the gas or protective gas for playing the role of being spaced vaporization prevention phase and microlayer model；
(b) to manipulate the liquid that probe carrying is constituted microlayer model, vaporization prevention is passed through mutually to enter in confined space, in drop plate
In the groove of surface or drop plate or in drop layer surface hydrophobe limited area, microlayer model is formed；
(c) other functional liquids are carried to manipulate probe, passes through vaporization prevention mutually to enter in confined space, by the functionality
Liquid timing is quantitatively added in microlayer model；Sequence carries out aforesaid operations, and microlayer model is added in various other functional liquids
In；The type of other functional liquids includes sample liquid or reaction solution or buffer solution or dilution or cell suspension or training
Nutrient solution or stimulation liquid or induction liquid or rupture of membranes agent or precipitating reagent or denaturant or enzymatic reagent or immunoreagent or other
It is one or more in functional solution；
(d) heat treatment is entered to the Microfluidic droplet device or refrigeration is handled, or be incubated (incubation) processing, to complete
Reaction or cell culture in microlayer model or other operations；Using the seal of the Microfluidic droplet device, ensure microlayer model body
Product is in above process without reducing or not significantly reducing；
(e) it to manipulate probe, passes through vaporization prevention mutually to enter in confined space, is contacted with microlayer model, by the portion in microlayer model
Liquid separation body or whole liquid are fetched into except Microfluidic droplet device, carry out subsequent detection, analysis or other operations；
(f) aforesaid operations can carry out flexible combination, or change operation order according to specific needs.
It is contacted with microlayer model preferably, passing through vaporization prevention mutually to enter in confined space using manipulation probe, removes above-mentioned liquid
It drips generation, liquid feeding and takes liquid operation outer, can also carry out the operations such as the migration, fusion, division of drop；Utilize multiple manipulation probe structures
At probe array, operation repetitive is carried out to the microlayer model array that multiple microlayer models are constituted.
Preferably, for manipulate probe and vaporization prevention cover board, spacing board, drop plate three combination, by the two it
One, be fixed on translation stage or the two respectively while being fixed on different translation stages, realize to microlayer model manipulates it is accurate calmly
Position；In the above operation, the automation for being accurately positioned with manipulating to microlayer model is realized using auto-translating platform.
Preferably, processing vaporization prevention cover board or spacing board or drop plate using transparent material so that entirely manipulating
In processing procedure, imaging or positioning or on-line real-time measuremen can be carried out to microlayer model.
According to the present invention, in confined space, microlayer model nearby adds and forms identical liquid with microlayer model solvent, but not
It is contacted with microlayer model, using its volatilization in confined space, further suppresses evaporation of the microlayer model solvent in confined space.
According to the present invention, in confined space, by microlayer model addition need to subsequently be added the various reagents of microlayer model, but not with
Microlayer model contacts, and can not only utilize its volatilization in confined space, further suppress microlayer model solvent in confined space
Evaporation；Also confined space is mutually left so that manipulating probe and not needing guiding through vaporization prevention, you can various reagents are added into microlayer model, prevent
It only manipulates probe surface when across vaporization prevention phase and is mutually polluted or interfered by vaporization prevention its normal operating.
According to the present invention, microlayer model in a short time without apparent evaporation under conditions of, or when having special requirement, anti-
Vaporization prevention phase can not be used in evaporation layer, manipulation probe can enter directly through vaporization prevention layer manipulates microlayer model in confined space,
To prevent the interference of pollution or the operation of vaporization prevention relative manipulation probe.
According to the present invention, because the vaporization prevention of liquid mutually has certain permeability, behaviour need not be utilized whithin a period of time
When vertical probe passes through vaporization prevention mutually to enter in confined space to manipulate microlayer model, or when there are other special requirements, solid material is utilized
Vaporization prevention cover board is completely closed by material, i.e., the confined air surrounded by vaporization prevention cover board (including vaporization prevention phase), spacing board, drop plate
Between realize and be completely closed, thoroughly eliminate the evaporation of microlayer model.
According to the present invention, by changing the type of vaporization prevention cover board, spacing board, the material of drop plate or vaporization prevention phase,
The evaporation rate of microlayer model internal solvent can be regulated and controled, carry out such as protein crystallization experiments；Or make device that there is certain breathe freely
Property, it can be used for cell culture related experiment, such as cell two dimension, dimensional culture, drug screening, cell interaction based on cell
Deng.
According to the present invention, the kind full of the gas for playing the role of being spaced vaporization prevention phase and microlayer model in the confined space
Class has the combination of air or carbon dioxide or nitrogen or inert gas or other gases or more than one gases.As
It is preferred that being conducive to prevent the component in drop from chemical change occurs using nitrogen or other inert gases.
According to the present invention, using with volatile liquid as vaporization prevention phase, such as water or aqueous solution.In operating process
In, new vaporization prevention phase need to be periodically added, to supplement the loss that vaporization prevention mutually itself is generated by evaporation.Preferably, selection compared with
The spatial volume of thick vaporization prevention layer thickness or larger carrying vaporization prevention phase, cocoa store enough vaporization prevention phases, reduce it
The frequency of addition.
Advantages of the present invention essentially consists in：
(1) microlayer model is mutually not directly contacted with using vaporization prevention, utilizes gas by the two in the local confined space of formation
The method at interval can avoid the cross-contamination issue between microlayer model and vaporization prevention phase, microlayer model and microlayer model completely, meanwhile,
The vaporization prevention and interval problem of microlayer model can be solved again；
(2) volume of microlayer model can be as small as 1 A Sheng, can effectively reduce sample and reagent consumption, realize to ultramicron
In the processing of sample, and avoid generating cross contamination；
(3) liquid vaporization prevention is used mutually to participate in the confined space of structure package microlayer model, you can the evaporation of microlayer model is prevented,
A variety of manipulations are carried out with respect to microlayer model so that manipulating probe and may pass through vaporization prevention again simultaneously, manipulation device is simple in structure, is easy to take
It builds, the flexibility of manipulation, simplicity and highly reliable, it is easy to accomplish the array of operation and automation；
It (4), can be with regulating device while the cross-contamination issue in solution drop experiment, microlayer model evaporation problems
Gas permeability, carry out drop in cell culture and subsequent operation；
(5) wide applicability can be applied to the biochemical analysis, high-flux medicaments sifting, albumen of ultramicron Sample volume
The screening of matter crystallization condition, cell analysis, even single cell analysis, single vesica analysis, single molecule analysis etc..
Description of the drawings
Fig. 1 is the Microfluidic droplet chip apparatus of the anti-drop evaporation of channel interval formula of the embodiment of the present invention 1, device
Vertical section schematic diagram.
Fig. 2 is the experimental result picture that the Microfluidic droplet chip apparatus of embodiment 1 is used to prevent drop from evaporating.
Fig. 3 a are a kind of contactless vaporization prevention Microfluidic droplets for structure that embodiment 2 is used to study cell drop culture
Chip apparatus is the vertical section schematic diagram of device.The device is mutually present in the situation in through-hole suitable for vaporization prevention.
Fig. 3 b are the contactless vaporization prevention miniflow drainings for another structure that embodiment 2 is used to study cell drop culture
Chip apparatus is dripped, is the vertical section schematic diagram of device.Spacing board is not used in the device, and vaporization prevention is mutually present in vaporization prevention cover board
In through-hole, microlayer model is located in the groove structure of drop plate.
Fig. 3 c are the contactless vaporization prevention miniflow drainings for the third structure that embodiment 2 is used to study cell drop culture
Chip apparatus is dripped, is the vertical section schematic diagram of device.Vaporization prevention is mutually present in vaporization prevention lid surface in the device.
Fig. 3 d are the contactless vaporization prevention miniflow drainings for the 4th kind of structure that embodiment 2 is used to study cell drop culture
Chip apparatus is dripped, is the vertical section schematic diagram of device.Through-hole structure on spacing board with holes is comb shaped structure.Prevent in the device
Evaporation is mutually present in through-hole structure.
Fig. 4 a are a kind of Microfluidic droplet chip dresses for structure that embodiment 3 can complete trace protein pretreatment work
It sets, is the vertical section schematic diagram of device.And Fig. 3 a, Fig. 3 d devices are applied to the device used when single microlayer model operates
Diagrammatic cross-section.Vaporization prevention is mutually present in vaporization prevention cover plate through hole in the device.
Fig. 4 b are another Microfluidic droplet chip apparatus that embodiment 3 can complete trace protein pretreatment work,
It is the vertical section schematic diagram of device.It is also the partial enlarged view of a microlayer model unit in Fig. 3 b.Interval is not used in the device
Plate, vaporization prevention are mutually present in vaporization prevention cover plate through hole.
Fig. 4 c are the third Microfluidic droplet chip apparatus that embodiment 3 can complete trace protein pretreatment work,
It is the vertical section schematic diagram of device；And the section of device that the device of Fig. 3 c is applied to use when single microlayer model operates shows
It is intended to.
Fig. 4 d be Fig. 3 d device in a microlayer model unit partial side view.
Fig. 5 a are the schematic top plan views of Fig. 3 a.
Fig. 5 b are the schematic top plan views of Fig. 3 b.
Fig. 5 c are the schematic top plan views of Fig. 3 c.
Fig. 5 d are the schematic top plan views of Fig. 3 d.
Fig. 6 is microlayer model generation and the operation chart by taking Fig. 4 a devices as an example.
In figure：1- microlayer models, 2- vaporization prevention phases, 3- vaporization prevention cover boards, 4- spaced gas, 5- spacing boards, 6- drop plates, 7-
Manipulate probe, 8- confined spaces.
Specific implementation mode
With specific embodiment, the present invention will be further described below, and but the scope of the present invention is not limited thereto.
Embodiment 1
The Microfluidic droplet chip apparatus for preventing drop from evaporating is device longitudinal sectional view referring to Fig. 1, Fig. 1.Whole device
It is rounded, it is placed in the transparent plastic culture dish of 6 centimetres of diameter, chip apparatus lower end bottom surface is affixed with culture dish face.For holding
The vaporization prevention cover board 3 with through-hole for carrying vaporization prevention phase 2 is formed by acrylic board through rapid laser carving, and acrylic board thickness is
4.0 millimeters, vaporization prevention cover board 3 is equipped with through-hole, and through-hole diameter is 1.0 millimeters, and vaporization prevention is mutually carried in the through-hole.
Directly use culture dish bottom surface as the drop plate 6 of carrying microlayer model 1, thickness is 1.5 millimeters.Vaporization prevention cover board with holes
There is one layer of circular ring shape spacing board 5 between 3 and drop plate 6, be dimethyl silicone polymer (PDMS) material, thickness is 3.0 millimeters, is used
It is contacted with vaporization prevention phase 2 to avoid droplets from 1.Utilize epoxide-resin glue by two between vaporization prevention cover board 3 and angular interval plate 5 with holes
Person's contact surface closely binds, angular interval plate 5 and drop plate 6 --- it can reach close by directly fitting between culture dish bottom surface
Seal effect.And all microlayer models 1 can be surrounded, form closed system.
Confined space 8 is surrounded by vaporization prevention cover board 3, vaporization prevention phase 2, spacing board 5 and drop plate 6, is filled in confined space 8
There are spaced gas 4, spaced gas 4 that can select air or carbon dioxide or nitrogen or inert gas or other gases, or
The combination etc. of more than one gases of person, it is preferred to use nitrogen or other inert gases are conducive to prevent the component in drop
Learn variation.
For generating dyestuff microlayer model, all it is added into each through-hole of vaporization prevention cover board 3 with holes using pipettor
Mineral oil is as vaporization prevention phase 2.It recycles capillary probe as probe 7 is manipulated, passes through the mineral oil vaporization prevention phase in through-hole
2, in drop plate 6 --- the dyestuff microlayer model 1 that 300 nanoliters are formed on culture dish bottom surface sequentially generates different drops, forms liquid
Drip array.It places 1 day, 2 days, 3 days, drop size is observed using microscope, study the evaporation of droplet array in chip
Situation.
Fig. 2 is the result of study of dyestuff microlayer model array evaporation situation after placing different number of days.Show to make using mineral oil
For vaporization prevention phase 2, microlayer model 1 is almost without evaporation.The vaporization prevention effect of different vaporization prevention phases 2 is also inconsistent, and can utilize should
Device is studied under different vaporization prevention phases 2, the evaporation situation of microlayer model 1.
Embodiment 2
Regular growth culture is carried out in culture bottle or culture dish, and cell is pressed respective existence feature and given birth in a reservoir
Long, single layer apposition growth, suspension cell are in that suspended state is grown in the medium to attached cell on the surface.In conventional drop
Cell culture is then that the drop containing cell and its culture solution is wrapped in oil phase to cultivate.And the present invention is utilized, it can be with
So that cell and its culture solution is not contacted vaporization prevention phase, carry out drop inner cell culture, cell may mutually be trained to avoid vaporization prevention
Support the interference generated and adverse effect.
Study cell drop culture contactless vaporization prevention Microfluidic droplet chip apparatus, referring to Fig. 3 a, 3b, 3c and
3d, Fig. 3 a, 3b, 3c and 3d are the longitudinal sectional view of device.Fig. 4 a, 4b, 4c and 4d are respectively corresponding to above-mentioned 4 devices
Partial enlarged view or unit operations longitudinal sectional view, Fig. 5 a, 5b, 5c and 5d are respectively Fig. 3 a, 3b, 3c and 3d corresponding
The vertical view of device.The longitudinal sectional view of Fig. 3 d devices shows through-hole on vaporization prevention cover board 3 with holes or through-hole array in addition to small
Outside circle/square hole, the comb arrays of rectangular slat band and its formation are can also be, while can be found in Fig. 5 d.It is entire contactless anti-
It is rounded to evaporate Microfluidic droplet chip apparatus, is placed in the transparent plastic culture dish of 6 centimetres of diameter.For carrying vaporization prevention phase
2 vaporization prevention cover board 3 with holes is formed by PDMS plates through manual perforation processing, and PDMS plate thickness is 3.0 millimeters, and through-hole diameter is
1.0 millimeter.Drop plate 6 for carrying microlayer model 1 is PDMS material, and thickness is 3.0 millimeters.Vaporization prevention cover board 3 and liquid with holes
There are one layer of circular ring shape spacing board 5 and PDMS material between drop plate 6, thickness is 3.0 millimeters, for avoiding microlayer model 1 and anti-steaming
It sends out phase 2 to contact, and all microlayer models 1 can be surrounded, form closed system.And 3 top of vaporization prevention cover board with holes, it can also add one
The support component surrounded vaporization prevention phase 2, prevent it from overflowing, is equally to process to obtain using PDMS material.It is support component, with holes anti-
Evaporate cover board 3, spacing board 5 and drop plate 6, after all first passing through oxygen plasma treatment surface, complete each plane according to
It is secondary to fit closely.
It has used oil phase liquid and aqueous phase liquid as vaporization prevention phase 2 respectively, has carried out the culture of cell microlayer model 1；Oil phase
Liquid corresponding diagram 3a (Fig. 3 b and Fig. 3 d also can), aqueous phase liquid corresponding diagram 3c.
When oil phase liquid being selected to be fluorinated liquid FC-40 as vaporization prevention phase 2, i.e., tested using Fig. 3 a devices.Utilize shifting
FC-40 is all added as vaporization prevention phase 2 into each through-hole of vaporization prevention cover board 3 with holes in liquid device.It is manipulated again by capillary
Probe 7 passes through the vaporization prevention phase 2 in through-hole, forms 300 nanoliters of cell microlayer models 1 in the plane of drop plate 6, sequentially forms
Cell droplet array.Then culture dish is covered, whole device is put into cell incubator, to prevent vaporization prevention phase 2 from evaporating
It causes cell microlayer model 1 to evaporate soon, a small amount of water is added in chip apparatus periphery, evaporates measure as additional anti-drop.It puts
After setting 24 hours, probe 7 is manipulated by capillary, it is live/dead (live/dead) that prepared cell is added into cell drop 1
300 nanoliters of staining reagent.After being put into incubator dyeing half an hour, whole device is moved on to thin in fluorescence microscopy microscopic observation drop
Born of the same parents' staining conditions obtain the cell survival rate in drop by being counted respectively to living cells, dead cell.
Result of study is shown, after 24 hours, all evaporations of FC-40 vaporization preventions phase 2, but cell microlayer model 1 is due to additional
Measure, evaporation situation is not obvious, while cell survival rate reaches 50% or more.This shows before 2 evaporating completely of vaporization prevention phase,
Cells at least more than half are still survived.
It when using water as vaporization prevention phase 2, i.e., is tested using Fig. 3 c devices, this is because the characteristic of water, it can't
It is stable in the presence of in PDMS through-holes, but on the surfaces PDMS.Using pipettor, water is injected to 3 surface of vaporization prevention cover board with holes
As vaporization prevention phase 2, water layer covers all through-holes.Probe 7 is manipulated by capillary again, gently passes through the water in through-hole (anti-
Only water is stabbed into through-hole and is fallen), 300 nanoliters of cell microlayer models 1 are formed in the plane of drop plate 6, sequentially form cell liquid
Drip array.Then culture dish is covered, whole device is put into cell incubator.After placing 24 hours, manipulated by capillary
Live/dead 300 nanoliters of (live/dead) staining reagent of prepared cell is added into cell microlayer model 1 for probe 7.It is put into culture
Case dye half an hour after, whole device is moved on into fluorescence microscopy microscopic observation drop inner cell staining conditions, by living cells,
Dead cell counts respectively, obtains the cell survival rate in drop.
Result of study is shown, after 24 hours, water layer vaporization prevention phase 2 there is no evaporating completely, the cell microlayer model 1 of lower section
Evaporation situation is not obvious, and cell survival rate also can reach 60% or more.
The survival ability of different cells is inconsistent, it is possible to study different 1 inner cells of cell microlayer model using the device
Existing state, obtain the survival ability of different cells.The gas permeability of different vaporization prevention phases 2, bio-compatibility are also inconsistent, institute
Also using the device by the survival rate of different 2 times drops of vaporization prevention phase, 1 inner cells of observation, to obtain different vaporization prevention phases 2
Gas permeability or bio-compatibility.
Embodiment 3
The Microfluidic droplet chip apparatus that can complete trace protein sample pretreatment, referring to Fig. 4 a, 4b or 4c, this
It is the pretreatment unit for a sample.Multi-example, high-throughput pretreatment can be by integrating above-mentioned self-contained unit, i.e.,
Obtain Fig. 3 a, 3b or 3d.
Such as Fig. 4 a of device used in experiment.Whole device is rounded, the vaporization prevention lid with holes for carrying vaporization prevention phase 2
Plate 3 is formed by PDMS blocks through manual perforation processing, and PDMS layer thickness is 3.0 millimeters, and through-hole diameter is 1.3 millimeters.For carrying
The drop plate 6 of microlayer model 1 is glass material, and thickness is 3.0 millimeters.There is one layer of circle between vaporization prevention cover board 3 and drop plate 6 with holes
Annular space plate 5 and glass material, thickness are 2.0 millimeters, and through-hole diameter is 1.5 millimeters, can avoid microlayer model 1 and prevent
It evaporates phase 2 to contact, prevents the cross contamination between vaporization prevention phase 2 and microlayer model 1 from causing the loss of sample fraction in drop.It is with holes
Vaporization prevention cover board 3, spacing board 5 and 6 each plane of drop plate are fitted closely or are binded successively.
In conjunction with Fig. 6, using pipettor, into each through-hole of vaporization prevention cover board 3 with holes, mineral oil is all added as anti-steaming
Send out phase 2.Probe 7 is manipulated by capillary again, the mineral oil vaporization prevention phase 2 in through-hole is passed through, is formed in the plane of drop plate 6
100 nanoliters of protein sample microlayer models 1.Then by reaction requirement, probe 7 is manipulated using capillary, is first added in backward microlayer model 1
Enter protein denaturant, reducing agent, alkylating reagent, enzymolysis reagent and terminates enzymolysis reagent.
Experiment also using Fig. 3 a, 3b 3d devices, is carried out at the same time the protein pretreatment behaviour of various sample drop 1
Make.
Claims (10)
1. a kind of micro flow control chip device of the anti-drop evaporation of channel interval formula, which is characterized in that including：
Manipulation probe (7) for generating and manipulating microlayer model (1)；
Drop plate (6) for carrying microlayer model (1), the drop loading end of the drop plate (6), which is equipped with, is used to form microlayer model
One or more drop zones；
Vaporization prevention cover board (3) of the closed the cover on drop plate (6) drop loading end, liquid described in vaporization prevention cover board (3) face
The position in drop area is equipped with the through-hole structure for being passed through for manipulating probe (7) when generating or manipulating drop；The through-hole structure
Interior or through-hole structure top surface is sealed with vaporization prevention phase (2)；
Confined space is formed between the vaporization prevention cover board (3) and drop plate (6), gas is filled in confined space；And ensure
After microlayer model generates, there are external series gaps between the vaporization prevention phase (2) and microlayer model (1).
2. the micro flow control chip device of the anti-drop evaporation of channel interval formula according to claim 1, which is characterized in that described
Vaporization prevention cover board (3) realizes that sealing is fixed with drop plate (6) by spacing board (5).
3. the micro flow control chip device of the anti-drop evaporation of channel interval formula according to claim 1, which is characterized in that described
Drop zone is groove structure.
4. the micro flow control chip device of the anti-drop evaporation of channel interval formula according to claim 1, which is characterized in that described
Through-hole structure is mutually independent single hole arrangements of the setting in vaporization prevention cover board (3), in each single hole arrangements and the drop zone
Under to just；Either the through-hole structure is bar shaped pore structure or comb shape pore structure.
5. the micro flow control chip device of the anti-drop evaporation of channel interval formula according to claim 1, which is characterized in that described
It is filled in confined space one or more in air, carbon dioxide, nitrogen, inert gas.
6. the micro flow control chip device of the anti-drop evaporation of channel interval formula according to claim 1, which is characterized in that single
Drop zone periphery is equipped with spacing board (5), and spacing board (5) top and bottom are closed with vaporization prevention cover board (3) and drop plate (6) respectively
Connection, two neighboring drop zone are mutually isolated by the spacing board (5).
7. the micro flow control chip device of the anti-drop evaporation of channel interval formula according to claim 1, which is characterized in that described
The volume of microlayer model is that 1 Ah rises to 1 milliliter.
8. the micro flow control chip device of the anti-drop evaporation of channel interval formula according to claim 1, which is characterized in that described
Vaporization prevention phase (2) is mineral oil, dimethicone, fluorocarbon oil, water or aqueous solution.
9. a kind of micro flow control chip device using the anti-drop evaporation of any one of the claim 1-8 channel interval formulas carries out micro-
The method of flow-control operation, which is characterized in that including：Using the liquid for manipulating probe carrying composition microlayer model, vaporization prevention phase is passed through
Into in confined space, in the drop zone on drop plate (6) surface, microlayer model is formed；Or pass through anti-steaming using probe is manipulated
Hair mutually enters in confined space, liquid is added into the microlayer model formed, or by taking out liquid in microlayer model.
10. the method according to claim 9 for carrying out micro-fluidic operation, which is characterized in that in confined space, in micro- liquid
Drop addition nearby forms identical liquid with microlayer model solvent.
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| CN115178311A (en) * | 2022-06-08 | 2022-10-14 | 北京机械设备研究所 | Electrowetting drive chip for controlling various liquid drops and control method thereof |
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|---|---|---|---|---|
| US5180555A (en) * | 1988-02-16 | 1993-01-19 | Bio Merieux | Microbiological analysis cup or the like |
| CN201658976U (en) * | 2010-03-12 | 2010-12-01 | 胡家元 | Anti-pollution evaporation pan with cover |
| CN103290487A (en) * | 2012-02-29 | 2013-09-11 | 科美仪器 | Bio-reaction device chip |
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| JPS63200063A (en) * | 1987-02-16 | 1988-08-18 | Olympus Optical Co Ltd | Plate for measuring immunoreaction |
| US5789251A (en) * | 1994-06-16 | 1998-08-04 | Astle; Thomas W. | Multi-well bioassay tray with evaporation protection and method of use |
| DE10020771A1 (en) * | 2000-04-28 | 2001-10-31 | Merck Patent Gmbh | Pipetting device |
| FI20105591A0 (en) * | 2010-05-26 | 2010-05-26 | Arcdia Internat Oy Ltd | EXCLUSION OF REACTION CABLES FOR BIOAFFINITY ASSAYS |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5180555A (en) * | 1988-02-16 | 1993-01-19 | Bio Merieux | Microbiological analysis cup or the like |
| CN201658976U (en) * | 2010-03-12 | 2010-12-01 | 胡家元 | Anti-pollution evaporation pan with cover |
| CN103290487A (en) * | 2012-02-29 | 2013-09-11 | 科美仪器 | Bio-reaction device chip |
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