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 PDF

Info

Publication number
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
Authority
CN
China
Prior art keywords
pdms
paper base
array
single cell
cell analysis
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201811432712.5A
Other languages
Chinese (zh)
Other versions
CN109289954B (en
Inventor
孙浩
东辉
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Fuzhou University
Original Assignee
Fuzhou University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Fuzhou University filed Critical Fuzhou University
Priority to CN201811432712.5A priority Critical patent/CN109289954B/en
Publication of CN109289954A publication Critical patent/CN109289954A/en
Application granted granted Critical
Publication of CN109289954B publication Critical patent/CN109289954B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027Containers 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
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10Investigating individual particles
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/10Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/08Geometry, shape and general structure
    • B01L2300/0887Laminated structure
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/12Specific details about materials
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/12Specific details about materials
    • B01L2300/126Paper

Landscapes

  • 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)
  • Immunology (AREA)
  • 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

A kind of compound micro-fluidic chip of array PDMS- paper base for single cell analysis and Its control method
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.
CN201811432712.5A 2018-11-28 2018-11-28 Array PDMS-paper-based composite microfluidic chip for single cell analysis and control method thereof Active CN109289954B (en)

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

Applications Claiming Priority (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

Publications (2)

Publication Number Publication Date
CN109289954A true CN109289954A (en) 2019-02-01
CN109289954B CN109289954B (en) 2024-04-09

Family

ID=65143290

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201811432712.5A Active CN109289954B (en) 2018-11-28 2018-11-28 Array PDMS-paper-based composite microfluidic chip for single cell analysis and control method thereof

Country Status (1)

Country Link
CN (1) CN109289954B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
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

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104130932A (en) * 2014-07-30 2014-11-05 华中科技大学 Agarose microfluidic chip based bacteria enrichment device
CN106040326A (en) * 2016-05-27 2016-10-26 南京大学 Macro-porous paper based-polydimethylsiloxane composite microfluidic chip and preparation method and application thereof
WO2018071448A1 (en) * 2016-10-11 2018-04-19 The Regents Of The University Of California Systems and methods to encapsulate and preserve organic matter for analysis
CN108421566A (en) * 2018-05-18 2018-08-21 福州大学 A kind of paper substrate micro-fluidic chip array system and digitlization concurrency control method
CN209222159U (en) * 2018-11-28 2019-08-09 福州大学 A kind of compound micro-fluidic chip of array PDMS- paper base for single cell analysis

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104130932A (en) * 2014-07-30 2014-11-05 华中科技大学 Agarose microfluidic chip based bacteria enrichment device
CN106040326A (en) * 2016-05-27 2016-10-26 南京大学 Macro-porous paper based-polydimethylsiloxane composite microfluidic chip and preparation method and application thereof
WO2018071448A1 (en) * 2016-10-11 2018-04-19 The Regents Of The University Of California Systems and methods to encapsulate and preserve organic matter for analysis
CN108421566A (en) * 2018-05-18 2018-08-21 福州大学 A kind of paper substrate micro-fluidic chip array system and digitlization concurrency control method
CN209222159U (en) * 2018-11-28 2019-08-09 福州大学 A kind of compound micro-fluidic chip of array PDMS- paper base for single cell analysis

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
张国豪;马波;秦建华;林炳承;: "集成药物代谢微流控芯片的研制", 高等学校化学学报, no. 07 *

Cited By (1)

* Cited by examiner, † Cited by third party
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

Also Published As

Publication number Publication date
CN109289954B (en) 2024-04-09

Similar Documents

Publication Publication Date Title
CN209222159U (en) A kind of compound micro-fluidic chip of array PDMS- paper base for single cell analysis
US10927407B2 (en) Systems and methods for handling microfluidic droplets
US10639597B2 (en) Microfluidic devices
Kong et al. Lab-on-a-CD: A fully integrated molecular diagnostic system
Marshall et al. Integrated printed circuit board device for cell lysis and nucleic acid extraction
CN103103120A (en) Integrated temperature controlled PCR-CE (polymerase chain reaction-capillary electrophoresis) micro-fluidic chip and preparation method thereof
EP2981349A2 (en) Systems and methods for handling microfluidic droplets
CN108940387A (en) It is a kind of for unicellular isolated drop micro-fluidic chip and preparation method thereof
US20140255270A1 (en) Removing sacrificial layer to form liquid containment structure and methods of use thereof
CN108315389A (en) A kind of micro-volume cellular nucleic acid amplification method
CN205170857U (en) A micro -fluidic chip , detecting system and device for DNA short -term test
CN112076807A (en) Micro-fluidic chip and device capable of spontaneously forming water-in-oil droplets
CN209397220U (en) Micro-fluidic chip and capture drop carry out the device of nucleic acid amplification
CN109289954A (en) A kind of compound micro-fluidic chip of array PDMS- paper base and its control method for single cell analysis
Yin et al. Micro/Nanofluidic‐Enabled Biomedical Devices: Integration of Structural Design and Manufacturing
Furuberg et al. RNA amplification chip with parallel microchannels and droplet positioning using capillary valves
CN102162807B (en) Nucleic acid array analysis platform and method based on microfluidic chip
CN103894247B (en) A kind of nucleic acid multiplex amplification micro-fluidic chip
EP2840130B1 (en) Dna chip with micro-channel for dna analysis
JP2008017779A (en) Lab-on-a-chip
Li et al. Insights and Advancements in Microfluidics
Grabowska-Jadach et al. Lab-on-a-chip systems for biomedical analysis
Liou et al. Investigated of DNA microarray chips for disease detection and gene sequencing
Lv et al. Continue flow PCR-CE chip for DNA analysis
Madadi Analysis and design of a capillary driven blood plasma separation microfluidic device

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant