CN109289954A - A kind of compound micro-fluidic chip of array PDMS- paper base and its control method for single cell analysis - Google Patents
A kind of compound micro-fluidic chip of array PDMS- paper base and its control method for single cell analysis Download PDFInfo
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- CN109289954A CN109289954A CN201811432712.5A CN201811432712A CN109289954A CN 109289954 A CN109289954 A CN 109289954A CN 201811432712 A CN201811432712 A CN 201811432712A CN 109289954 A CN109289954 A CN 109289954A
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- 238000004458 analytical method Methods 0.000 title claims abstract description 27
- 150000001875 compounds Chemical class 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 title claims abstract description 18
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 claims abstract description 33
- 239000004205 dimethyl polysiloxane Substances 0.000 claims abstract description 31
- 235000013870 dimethyl polysiloxane Nutrition 0.000 claims abstract description 31
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 claims abstract description 31
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims abstract description 31
- 239000000758 substrate Substances 0.000 claims abstract description 17
- 239000011159 matrix material Substances 0.000 claims abstract description 9
- 230000002209 hydrophobic effect Effects 0.000 claims abstract description 8
- 239000000463 material Substances 0.000 claims abstract description 8
- 210000004027 cell Anatomy 0.000 claims description 44
- 229920000936 Agarose Polymers 0.000 claims description 12
- 238000003752 polymerase chain reaction Methods 0.000 claims description 10
- 239000000126 substance Substances 0.000 claims description 8
- 238000010438 heat treatment Methods 0.000 claims description 7
- 230000003321 amplification Effects 0.000 claims description 6
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 6
- 238000005336 cracking Methods 0.000 claims description 5
- 239000011324 bead Substances 0.000 claims description 4
- 210000002421 cell wall Anatomy 0.000 claims description 4
- 125000004122 cyclic group Chemical group 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 239000000155 melt Substances 0.000 claims description 4
- 238000000197 pyrolysis Methods 0.000 claims description 4
- 239000011347 resin Substances 0.000 claims description 4
- 229920005989 resin Polymers 0.000 claims description 4
- 239000007790 solid phase Substances 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 3
- 239000000835 fiber Substances 0.000 abstract description 5
- 229920000642 polymer Polymers 0.000 abstract description 3
- 238000010586 diagram Methods 0.000 description 6
- 238000013461 design Methods 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 4
- 238000001514 detection method Methods 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 239000003814 drug Substances 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 238000001259 photo etching Methods 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 238000007397 LAMP assay Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 238000010195 expression analysis Methods 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- UQEAIHBTYFGYIE-UHFFFAOYSA-N hexamethyldisiloxane Polymers C[Si](C)(C)O[Si](C)(C)C UQEAIHBTYFGYIE-UHFFFAOYSA-N 0.000 description 1
- 238000010185 immunofluorescence analysis Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 229920005573 silicon-containing polymer Polymers 0.000 description 1
- 238000002174 soft lithography Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
Classifications
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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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
-
- 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
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
-
- 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
-
- 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
- B01L2300/12—Specific details about materials
-
- 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
- B01L2300/12—Specific details about materials
- B01L2300/126—Paper
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Dispersion Chemistry (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Clinical Laboratory Science (AREA)
- Hematology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
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- Pathology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention relates to a kind of compound micro-fluidic chips of array PDMS- paper base and its control method for single cell analysis, are disposed with PDMS layer, glue-line, paper substrate and heat patch from top to bottom；The capture array being made of more than one concave structure is provided in the PDMS layer；The paper substrate includes the array being made of the hydrophilic matrix unit including hydrophilic paper base structure and hydrophobic structure；Each concave structure each of in the paper substrate in hydrophilic matrix unit and the PDMS layer corresponds.The present invention can be used for single cell analysis, and chip takes into account two kinds of material advantages of PDMS polymer and filter paper fibre.
Description
Technical field
The present invention relates to micro-fluidic chip design field, especially a kind of array PDMS- paper for single cell analysis
The compound micro-fluidic chip of base and its control method.
Background technique
Dimethyl silicone polymer (Polydimethylsiloxane, PDMS) has good optical property, thermal stability
And the advantages that bio-compatibility, it is one of main rapidoprint of micro-fluidic chip.Chip based on PDMS material, in cell
Manipulation, genes within cells detection of expression and immunofluorescence analysis etc. successful application.But the fluid stream in the microchannel PDMS
Dynamic to usually require to rely on external syringe pump or starting micro-valve structure control, there is still a need for complete in the lab for analysis test.
In the past 10 years, the chip processed using filter paper as basis material, i.e. paper substrate micro-fluidic chip gradually become low cost,
Portable and environment-friendly type biochemistry detection tool.By intrinsic capillary fiber structure inside filter paper, in paper substrate micro-fluidic chip
Fluid driving and control are realized by capillarity, eliminate dependence of the conventional microfluidic control to equipment such as precise injection pumps, especially suitable
Biochemical sample detection and analysis should be carried out under the conditions of resource constraint.But because filter paper fibre structure is mixed and disorderly point usually random
Cloth, this structure are difficult to realize the precise manipulation of the low abundance sample (such as unicellular) of trace.
Summary of the invention
In view of this, the purpose of the present invention is to propose to a kind of array PDMS- paper base for single cell analysis is compound micro-
Fluidic chip and its control method can be used for single cell analysis, and chip takes into account PDMS polymer and two kinds of materials of filter paper fibre are excellent
Point.
The present invention is realized using following scheme: a kind of array PDMS- paper base for single cell analysis is compound micro-fluidic
Chip is disposed with PDMS layer, glue-line, paper substrate and heat patch from top to bottom；
The capture array being made of more than one concave structure is provided in the PDMS layer；
The paper substrate includes the array being made of the hydrophilic matrix unit including hydrophilic paper base structure and hydrophobic structure；
Each concave structure each of in the paper substrate in hydrophilic matrix unit and the PDMS layer corresponds.
Further, including the glue-line is low melting-point agarose.
Further, the fusing point of the low melting-point agarose is 65 DEG C.
Further, the material of the hydrophobic structure is photosensitive resin.
Further, the compound micro-fluidic chip periphery of the array PDMS- paper base for single cell analysis is using pressure
The encapsulation of power film.
It is compound micro- based on the array PDMS- paper base described above for single cell analysis that the present invention also provides a kind of
The control method of fluidic chip, specifically:
When cell current-carrying flows through the concave structure of PDMS layer, the cell of special diameter size will be detained in this configuration；
The PDMS layer and paper substrate are bonded by the glue-line that low melting-point agarose is constituted, and the heating of chip bottom is arranged in
The glue-line melts after patch heating, falls into hydrophilic paper base structure by cold and hot convection current cell；
When cell pyrolysis liquid flows through hydrophilic paper base structure, the cell wall chemical cracking of cell discharges including RNA, DNA
Substance, DNA molecular chain are extracted by the micro- magnetic bead of solid phase, and by polymerase chain reaction cyclic amplification, polymerase chain reaction is produced
Object signal is directly read by fluorescence microscope.
Compared with prior art, the invention has the following beneficial effects:
1, chip of the invention takes into account two kinds of material advantages of PDMS polymer and filter paper fibre, and microfluidic methods integrating cell is caught
It catches, fix, DNA extraction and PCR cycle amplification etc. are multi-functional, a variety of instrument and equipments required for previous cell experiment are eliminated,
Greatly simplify analysis experiment flow.
2, design of the invention has the multiple advantages such as easy to operate, low in cost and multipurpose, has innovated existing miniflow
Cell analysis method is controlled, to Personalized medicine platform is improved, promotes Portable Clinical diagnosis and treatment and the poor regional medical diagnosis on disease of substance
Deng with important potential using value.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of the embodiment of the present invention, wherein (a) is that schematic diagram is cutd open in side, it (b) is corresponding crosscutting signal
Figure.
Fig. 2 is the chip manufacturing flow diagram of the embodiment of the present invention.
Fig. 3 is the chip operation flow diagram of the embodiment of the present invention.
Specific embodiment
The present invention will be further described with reference to the accompanying drawings and embodiments.
It is noted that described further below be all exemplary, it is intended to provide further instruction to the application.Unless another
It indicates, all technical and scientific terms used herein has usual with the application person of an ordinary skill in the technical field
The identical meanings of understanding.
It should be noted that term used herein above is merely to describe specific embodiment, and be not intended to restricted root
According to the illustrative embodiments of the application.As used herein, unless the context clearly indicates otherwise, otherwise singular
Also it is intended to include plural form, additionally, it should be understood that, when in the present specification using term "comprising" and/or " packet
Include " when, indicate existing characteristics, step, operation, device, component and/or their combination.
As shown in Figure 1, it is compound micro-fluidic to present embodiments provide a kind of array PDMS- paper base for single cell analysis
Chip is disposed with PDMS layer, glue-line, paper substrate and heat patch from top to bottom；
The capture array being made of more than one concave structure is provided in the PDMS layer；
The paper substrate includes the array being made of the hydrophilic matrix unit including hydrophilic paper base structure and hydrophobic structure；
Each concave structure each of in the paper substrate in hydrophilic matrix unit and the PDMS layer corresponds.
In the present embodiment, including the glue-line is low melting-point agarose.
In the present embodiment, the fusing point of the low melting-point agarose is 65 DEG C.
In the present embodiment, the material of the hydrophobic structure is photosensitive resin.
In the present embodiment, the compound micro-fluidic chip periphery of array PDMS- paper base for single cell analysis is adopted
It is encapsulated with pressure membrane.
The present embodiment additionally provides a kind of compound based on the array PDMS- paper base described above for single cell analysis
The control method of micro-fluidic chip, specifically:
When cell current-carrying flows through the concave structure of PDMS layer, the cell of special diameter size will be detained in this configuration；
The PDMS layer and paper substrate are bonded by the glue-line that low melting-point agarose is constituted, and the heating of chip bottom is arranged in
The glue-line melts after patch heating, falls into hydrophilic paper base structure by cold and hot convection current cell；
When cell pyrolysis liquid flows through hydrophilic paper base structure, the cell wall chemical cracking of cell discharges including RNA, DNA
Substance, DNA molecular chain are extracted by the micro- magnetic bead of solid phase, and by polymerase chain reaction cyclic amplification, polymerase chain reaction is produced
Object signal is directly read by fluorescence microscope.
Preferably, the chip of the present embodiment design uses function Integration Design, there is unicellular capture, release, fixed point
The functions such as cracking, targeting DNA fragmentation amplification.The part PDMS is used for unicellular capture, using " recessed " shape three-dimensional structure, when cell carries
When stream flows through the structure, the cell of special diameter size will be detained in this configuration；To avoid flow dead zone and irregular convection,
" recessed " shape three-dimensional structure upper dimension is wide as shown in Figure 1 with bottom.The hydrophobic structure of paper base part is constructed by photosensitive resin, is passed through
The processing of Conventional UV photoetching process；Hydrophilic-structure after processing is in array distribution, each unit and PDMS " recessed " shape three-dimensional structure
It corresponds.It is bonded with paper base part by low melting-point agarose in the part PDMS, agar melts after heating, by cold and hot convection current
Cell is fallen into hydrophilic paper base.When cell pyrolysis liquid flows through paper base unit, cell wall chemical cracking discharges RNA, DNA etc..DNA points
Subchain is extracted by the micro- magnetic bead of solid phase, and passes through polymerase chain reaction (PCR) cyclic amplification.In chip bottom, temperature control patch is set
(heat patch) to real-time control and monitors temperature in each reaction member.To avoid the sample being likely to occur in reaction evaporation,
Chip periphery is encapsulated using pressure membrane.Because PDMS translucency is preferable, PCR reaction product signal can directly be read by fluorescence microscope
It takes.Entire cell analysis process, which does not need the equipment such as syringe pump, can be completed.The present embodiment be put forward for the first time for Manipulation of single cells,
The compound microfluidic methods of array PDMS- paper base and feasible chip Prototype Design of intracellular DNA analysis, have multifunctional unit
The advantages that change, has a good application prospect in terms of portable analysis, Personalized medicine and trace sample.
Fig. 1 is the compound microfluidic chip structure schematic diagram of the present embodiment array PDMS- paper base.It is set in the cavity of the part PDMS
" recessed " shape three-dimensional matrix structure is set, Cell capture and fixation are used for；Low melting-point agarose is for being isolated PDMS and filter paper part；Filter
Paper part is made of hydrophilic rectangular element array, and each unit is corresponding with PDMS " recessed " shape three-dimensional structure, is used for intracellular DNA temperature
Control amplified reaction.
Fig. 2 is the micro-fluidic chip PDMS structure schematic diagram of the process of the embodiment of the present invention, the processing master of PDMS structure
To be based on standard soft lithography process.To obtain not wide " recessed " shape three-dimensional structure, using secondary photoetching process, narrow base Duan Weizhong
Hollow structure is used to support top capturing structure and avoids flow dead zone to greatest extent.Fusing point is used to do bottom for 65 DEG C of agarose
Portion's sealing, PDMS is connected with paper base segment fluid flow/and closure realized by temperature control.
Fig. 3 is the micro-fluidic operational flowchart of PDMS- paper base compound chip of the present embodiment.Multiple steps can be compound in monolithic
It is completed on chip.To be easier to the reagent evaporation problems occurred when avoiding high temperature (95 DEG C), the present embodiment uses ring mediated isothermal side
Method (loop-mediated isothermal amplification) expands DNA sample, and this method reaction temperature is set as
65℃。
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
Claims (6)
1. a kind of compound micro-fluidic chip of array PDMS- paper base for single cell analysis, it is characterised in that: from top to bottom according to
It is secondary to be provided with PDMS layer, glue-line, paper substrate and heat patch；
The capture array being made of more than one concave structure is provided in the PDMS layer；
The paper substrate includes the array being made of the hydrophilic matrix unit including hydrophilic paper base structure and hydrophobic structure；
Each concave structure each of in the paper substrate in hydrophilic matrix unit and the PDMS layer corresponds.
2. a kind of compound micro-fluidic chip of array PDMS- paper base for single cell analysis according to claim 1,
It is characterized in that: being low melting-point agarose including the glue-line.
3. a kind of compound micro-fluidic chip of array PDMS- paper base for single cell analysis according to claim 2,
Be characterized in that: the fusing point of the low melting-point agarose is 65 DEG C.
4. a kind of compound micro-fluidic chip of array PDMS- paper base for single cell analysis according to claim 1,
Be characterized in that: the material of the hydrophobic structure is photosensitive resin.
5. a kind of compound micro-fluidic chip of array PDMS- paper base for single cell analysis according to claim 1,
Be characterized in that: the compound micro-fluidic chip periphery of array PDMS- paper base for single cell analysis is encapsulated using pressure membrane.
6. a kind of based on the described in any item compound miniflows of array PDMS- paper base for single cell analysis of claim 1-5
Control the control method of chip, it is characterised in that:
When cell current-carrying flows through the concave structure of PDMS layer, the cell of special diameter size will be detained in this configuration；
The PDMS layer and paper substrate are bonded by the glue-line that low melting-point agarose is constituted, and the heating of chip bottom is arranged in
The glue-line melts after patch heating, falls into hydrophilic paper base structure by cold and hot convection current cell；
When cell pyrolysis liquid flows through hydrophilic paper base structure, the cell wall chemical cracking of cell discharges including RNA, DNA
Substance, DNA molecular chain are extracted by the micro- magnetic bead of solid phase, and by polymerase chain reaction cyclic amplification, polymerase chain reaction is produced
Object signal is directly read by fluorescence microscope.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811432712.5A CN109289954B (en) | 2018-11-28 | 2018-11-28 | Array PDMS-paper-based composite microfluidic chip for single cell analysis and control method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811432712.5A CN109289954B (en) | 2018-11-28 | 2018-11-28 | Array PDMS-paper-based composite microfluidic chip for single cell analysis and control method thereof |
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| Publication Number | Publication Date |
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| CN109289954A true CN109289954A (en) | 2019-02-01 |
| CN109289954B CN109289954B (en) | 2024-04-09 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113713868A (en) * | 2021-09-13 | 2021-11-30 | 北京京东方技术开发有限公司 | Microfluidic chip and manufacturing method thereof |
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