CN201770704U

CN201770704U - Microfluidic chip for polymerase chain reaction (PCR)

Info

Publication number: CN201770704U
Application number: CN2010200331340U
Authority: CN
Inventors: 隋国栋; 刘超; 刘思秀; 张金玲; 赵望
Original assignee: Fudan University
Current assignee: Fudan University
Priority date: 2010-01-15
Filing date: 2010-01-15
Publication date: 2011-03-23
Anticipated expiration: 2020-01-15

Abstract

The utility model belongs to the field of biomedicine, relates to a microfluidic chip for a polymerase chain reaction (PCR). The chip comprises a substrate and indium tin oxide (ITO) conductive glass, wherein the substrate of a micro flow path unit is provided with a liquid inflow and outflow through hole, is realized by a soft photoetching method and has a micro flow path structure; three heating zones and two heat insulation zones are etched on the ITO conductive glass; the side face of each heating zone is provided with a pin connected with an external temperature controller; and a temperature sensor embedded in the microgroove of the substrate is connected with the pin. In the microfluidic chip, the defect existing in the conventional PCR amplification instrument is overcome, and a completely transparent temperature-controllable polydimethylsiloxane (PDMS) microfluidic chip is manufactured by optically transparent PDMS and ITO conductive glass. By the microfluidic chip, reaction speed can be greatly improved, the PCR can be realized rapidly only within 10 to 20 minutes, and flow speed can be changed and the choice range of sample capacity can be widened through an injection pump.

Description

A kind of micro-fluidic chip that is used for PCR

Technical field

The utility model belongs to biomedical sector, relates to the micro-fluidic chip system structure design, is specifically related to the micro-fluidic chip of a kind of PCR of can be used for.The utility model is mainly used in a plurality of fields such as scientific research, clinical diagnosis, immunology and animal and plant.

Background technology

(polymerase chain reaction PCR) because of it has represented unprecedented sensitivity and specificity, is subjected to people's great attention, has become the gordian technique of gene studies, is widely used in bio-science every field [1,2] in the polymerase chain reaction.It is a kind of to specifying gene fragment to realize the method for amplification that prior art discloses PCR, and its ultimate principle is that by the temperature of the outer synthesis system of control volume, impelling double-stranded DNA to unwind becomes single stranded DNA according to the semiconservative DNA replication mechanism in the cells in vivo division.Single stranded DNA and the artificial primer renaturation of formulating, under certain dNTP concentration, resistant to elevated temperatures archaeal dna polymerase extends along the strand masterplate according to primer location and becomes double-stranded DNA subsequently.High-temperature denatured, low temperature renaturation and thermophilic extend the three-step reaction circulation and carry out, and make target DNA be able to rapid amplification.

In the making of pcr amplification equipment, most important for temperature controlling.The PCR thermal cycler is the pcr amplification instrument that is widely used at present.Its mode of operation is that the reaction vessel in that micro-processor controlled temperature cycle is wherein inserted amplification mixture adds template or sample DNA, carries out temperature cycle 20-30 time, can finish amplification procedure.

Conventional pcr amplification instrument exists that thermal capacitance is big, rate of heating and speed of cooling is slow, sample consumes high shortcoming.Usually finishing an amplification cycles needs a few to tens of minutes usually, therefore need spend several hours for 24 round-robin amplification procedures, and big for the demand of reactant.So along with the development of the widespread use of round pcr and MEMS technology, micro-fluidic chip technology, the development of micro pcr chip has obtained increasing concern and development.But the micro pcr chip has, and lot of advantages for example can reduce thermal capacity, shortens amplification and analysis time, the raising analysis speed, for the consumption of sample little and cheap cost and disposability.

Summary of the invention

The purpose of this utility model is the deficiency at the conventional pcr amplification instrument of prior art, and the micro-fluidic chip of a kind of PCR of being used for is provided.The utility model has designed a kind of application encapsulating structure of PCR micro-fluidic chip of continuously streamed, can realize thermal cycling fast, simplifies experimental installation, improves amplification rate and amplification efficiency.

The utility model adopts has optically transparent PDMS and ITO conductive glass, has designed and produced the bright PDMS micro-fluidic chip of Controllable Temperature full impregnated.Make the warm area of sample flow through three local steady temperatures of difference.This mode can improve speed of response greatly, only needs tens minutes to realize the fast PCR reaction, also can change the range of choice that flow velocity has also enlarged sample capacity by syringe pump in addition.

Particularly, the utility model provides the micro-fluidic chip of a kind of PCR of can be used for, it is characterized in that, it is made up of the via hole (1) that links to each other with outside syringe pump, micro-fluidic unit (3) and the ITO conductive glass (7) that links to each other with outside temperature controller; The substrate (4) of the microfluidic circuit unit (3) that the usefulness soft lithographic method of the via hole (1) that is used for incident and outgoing liquid of leaving this chip realizes and be etched with three heating zones (5) and the ITO conductive glass (7) of two heat insulating belts (6) with microfluidic circuit structure (2), the pin (8) that links to each other with outside temperature controller is left in the side of each heating zone (5), and the interior temperature sensor (10) of microflute (9) that is embedded in substrate (4) links to each other with pin (8).

Temperature sensor described in the utility model (10) is embedded in the microflute (9) of substrate (4).

In the utility model, described three heating zones (5) of PCR thermal cycling that are used for are by realizing at ITO conductive glass (7) pattern etching.

In the utility model, described have the described substrate that microfluidic circuit unit (3) arranged and electroconductive ITO glass (7) is realized the bright PCR micro-fluidic chip of full impregnated by thermal bonding.

In the utility model, described each heating zone (5) on ITO conductive glass (7) is gone up by pin (8) and outside temperature controller and is heated.

In the utility model, the material of described micro-fluidic chip can be silicon, quartz, glass, plastics, as polymethylmethacrylate (PMMA), polycarbonate (PC), silica gel such as polydimethylsiloxane (PDMS) etc., the chip channel internal surface can be to modify or unmodified.

In the utility model, microfluidic circuit structure (2) adopts winding shape continuously streamed pcr chip configuration.

The working process that the utility model is used for the micro-fluidic chip of PCR is: by outside temperature controller three heating zones (5) on the temperature conditioning unit are set at three required different temperature of PCR reaction respectively, thereby make the microfluidic circuit part that it contacted be in different warm areas respectively.By outside syringe pump with the PCR reactant by being injected into microfluidic circuit unit (3) by via hole (1) fixed kapillary (11), and in microfluidic circuit structure (11), flowing under the driving of syringe pump, passed through different warm areas along with the PCR reactant is mobile and corresponding in microfluidic circuit unit (3), realize temperature cycle, finished pcr amplification.

The utility model has been simplified the PCR reaction unit, only needs this structure is linked to each other with outside temperature controller, and links to each other with syringe pump, can carry out the PCR reaction.And can realize being evenly distributed of separate and each warm area inside that three warm areas are seen.

For the ease of understanding, below will describe in detail of the present utility model by concrete drawings and Examples.It needs to be noted, specific examples and accompanying drawing only are in order to illustrate, obviously those of ordinary skill in the art can illustrate according to this paper, in scope of the present utility model the utility model is made various corrections and change, these corrections and change are also included in the scope of the present utility model.

Description of drawings:

Fig. 1: the utility model structural representation;

Fig. 2: the utility model structure upward view;

Fig. 3: the utility model structure vertical view;

Fig. 4: the utility model microfluidic circuit unit substrate upward view;

Number in the figure: 1, via hole; 2, microfluidic circuit structure; 3, microfluidic circuit unit; 4, substrate; 5, heating zone; 6, heat insulating belt; 7, ITO conductive glass; 8, pin; 9, microflute; 10, transmitter; 11, kapillary.

Embodiment

Embodiment 1

The structure of exemplary micro-fluidic chip as shown in Figure 1, it mainly comprises the via hole (1) that links to each other with outside syringe pump, micro-fluidic unit (3) from top to bottom and the ITO conductive glass (7) that links to each other with outside temperature controller is formed.Syringe pump has adopted hand gun commonly used in the present embodiment, architecture adopts the bright PDMS material of full impregnated to make, its whole size is the high 7.8mm of the wide 39.4mm of long 35mm, and wherein the thickness of PDMS substrate (4) is 5.6mm, and the thickness of ITO conductive glass (7) is 2.2mm.

Make with respective outer diameters be that the kapillary (11) of 500um inserts in the via hole (1), make this chip link to each other with outside syringe pump.The microfluidic circuit loop structure (2) that carries out the PCR reaction that has the soft lithographic method to realize on the substrate (2) of micro-fluidic unit (3), the number of times of pcr amplification is by the cycle index decision of modular construction in microfluidic circuit unit (3).Fig. 4 is 24 loop structures.

ITO conductive glass (7) transmittance is greater than 90%, and side's resistance is 14 Europe.Three heating zones (5) and two heat insulating belts (6) form with the hydrochloric acid pattern etching of dilution.Two heat insulating belts (6) width is 5mm.3 microflutes (9) at substrate (2) are embedded with temperature sensor (10), connect the temperature that obtains three warm areas in the micro-fluidic unit (3) in the substrate (2) by temperature controller.

At first need 5 minutes warm up time during work, make three warm areas in the micro-fluidic unit (3) reach the temperature value of setting by temperature controller respectively by last heating zone (5) of ITO conductive glass (7) and temperature sensor (10), the PCR reactant flows into micro-fluidic unit (3) by kapillary (11) after the preheating under the driving of syringe pump, and it is mobile along microfluidic circuit structure (2), along with microfluidic circuit unit (3) go up the residing warm area difference of different piece, the PCR reactant is also corresponding to have passed through three different warm areas, and circulation repeatedly, realize pcr amplification reaction.

Claims

1. a micro-fluidic chip that is used for PCR is characterized in that, it is made up of the via hole (1) that links to each other with outside syringe pump, micro-fluidic unit (3) and the ITO conductive glass (7) that links to each other with outside temperature controller; The substrate (4) of the microfluidic circuit unit (3) that the usefulness soft lithographic method of the via hole (1) that is used for incident and outgoing liquid of leaving this chip realizes and be etched with three heating zones (5) and the ITO conductive glass (7) of two heat insulating belts (6) with microfluidic circuit structure (2), the pin (8) that links to each other with outside temperature controller is left in the side of each heating zone (5), and the interior temperature sensor (10) of microflute (9) that is embedded in substrate (4) links to each other with pin (8).

2. the micro-fluidic chip that is used for PCR according to claim 1 is characterized in that: described temperature sensor (10) is embedded in the microflute (9) of substrate (4).

3. the micro-fluidic chip that is used for PCR according to claim 1 is characterized in that: described three heating zones (5) are used for the PCR thermal cycling, by realizing at ITO conductive glass (7) pattern etching.

4. the micro-fluidic chip that is used for PCR according to claim 1 is characterized in that: the substrate of described microfluidic circuit unit (3) is made by PDMS; The bright electroconductive ITO glass (7) of the substrate of the microfluidic circuit unit (3) that described full impregnated is bright and full impregnated is realized the bright PCR micro-fluidic chip of full impregnated by thermal bonding.

5. the micro-fluidic chip that is used for PCR according to claim 1 is characterized in that: each heating zone (5) on ITO conductive glass (7) is gone up by pin (8) and outside temperature controller and is heated.

6. the micro-fluidic chip that is used for PCR according to claim 1 is characterized in that: the material of described micro-fluidic chip is selected from silicon, quartz, glass or plastics.

7. the micro-fluidic chip that is used for PCR according to claim 6 is characterized in that: the material of described micro-fluidic chip is selected from polymethylmethacrylate, polycarbonate or polydimethylsiloxane.

8. the micro-fluidic chip that is used for PCR according to claim 1 is characterized in that: described chip channel internal surface is modified or unmodified.

9. the micro-fluidic chip that is used for PCR according to claim 1 is characterized in that: described microfluidic circuit structure (2) adopts winding shape continuously streamed pcr chip configuration.