CN113862140A - Portable liquid drop digital PCR device - Google Patents
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- CN113862140A CN113862140A CN202111136369.1A CN202111136369A CN113862140A CN 113862140 A CN113862140 A CN 113862140A CN 202111136369 A CN202111136369 A CN 202111136369A CN 113862140 A CN113862140 A CN 113862140A
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- 238000001917 fluorescence detection Methods 0.000 claims description 11
- 238000011304 droplet digital PCR Methods 0.000 claims description 10
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- 229920009441 perflouroethylene propylene Polymers 0.000 claims description 4
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- 229920001721 polyimide Polymers 0.000 claims description 4
- -1 polytetrafluoroethylene Polymers 0.000 claims description 4
- 229920001343 polytetrafluoroethylene Polymers 0.000 claims description 4
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6851—Quantitative amplification
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Abstract
The invention discloses a portable liquid drop digital PCR device, which comprises a substrate, a movable sliding block, an oil phase injector, a water phase injector, a constant force spring, an injector fixing bracket, a connecting capillary tube, a liquid drop generating module, a PCR amplification capillary tube, a first electric heating film and a second electric heating film, the device comprises an injector fixing support, a movable sliding block, an oil-phase injector, a water-phase injector and a constant force spring, wherein the injector fixing support is fixed at one end of a substrate, the movable sliding block is arranged at the other end of the substrate, the injector fixing support is respectively connected with one ends of the oil-phase injector, the water-phase injector and the constant force spring, the movable sliding block is respectively connected with the other ends of the oil-phase injector, the water-phase injector and the constant force spring, one end of a connecting capillary tube is respectively connected with the oil-phase injector and the water-phase injector, the other end of the connecting capillary tube is connected with a liquid drop generation module, the output end of the liquid drop generation module is a PCR amplification capillary tube, and a first electric heating film and a second electric heating film wrap the PCR amplification capillary tube respectively. The invention can be widely applied to the technical field of polymerase chain reaction devices.
Description
Technical Field
The invention relates to the technical field of polymerase chain reaction devices, in particular to a portable liquid drop digital PCR device.
Background
Droplet digital polymerase chain reaction (ddPCR) is a method for separating PCR samples into water-in-oil droplets by using microfluidic droplet technology, and can be used for absolute quantification of nucleic acids in the samples. The injection module in the existing PCR device mostly adopts a mechanical pump or a pneumatic pump to drive the water phase and the oil phase, and the module has larger volume and high cost; one side of the amplification pipeline is heated in a heating table, a heat conduction module or a heating sheet mode in the amplification module, and the temperature of one side close to a heat source is higher than that of the other side, so that the pipeline is heated unevenly. In addition, one side of the pipeline of the top low-temperature area is provided with a heat conduction module, the other side of the pipeline of the top low-temperature area is exposed in the air, and a large amount of heat can be taken away by air convection, so that the stability of the low-temperature area is poor, and the amplification efficiency is obviously influenced by the ambient temperature. In the existing device, no matter a single heat source or a multi-heat source mode, a heating table or a heating plate with large volume is needed, and portability and light weight cannot be realized.
Disclosure of Invention
In view of this, the embodiment of the present invention provides a simple and practical portable droplet digital PCR device to achieve portability of the device.
On one hand, the invention provides a portable liquid drop digital PCR device, which comprises a substrate, a movable sliding block, an oil phase injector, a water phase injector, a constant force spring, an injector fixing bracket, a connecting capillary tube, a liquid drop generating module, a first electrothermal film and a second electrothermal film, the liquid drop generation module comprises a PCR amplification capillary tube, an injector fixing support is fixed at one end of a substrate, a movable sliding block is arranged at the other end of the substrate, the injector fixing support is respectively connected with one ends of an oil phase injector, a water phase injector and a constant force spring, the movable sliding block is respectively connected with the other ends of the oil phase injector, the water phase injector and the constant force spring, one end of a connecting capillary tube is respectively connected with the oil phase injector and the water phase injector, the other end of the connecting capillary tube is connected with the liquid drop generation module, the output end of the liquid drop generation module is the PCR amplification capillary tube, and a first electric heating film and a second electric heating film respectively wrap the PCR amplification capillary tube.
Optionally, the device further comprises a digital display temperature control platform, and the digital display temperature control platform is respectively connected with the first electric heating film and the second electric heating film and used for controlling the temperature of the first electric heating film and the temperature of the second electric heating film.
Optionally, the device further comprises a fluorescence detection module, wherein the fluorescence detection module comprises a microscope camera, a filter and a computer, the microscope camera is used for shooting and recording the state of the liquid drops flowing through the observation area, the filter is used for filtering light except the fluorescent probe in the sample, and the computer is used for detecting the fluorescence intensity of each liquid drop passing through the observation area and counting the number of the liquid drops passing through the observation area.
Optionally, the temperature of the first electro-thermal film is set to ninety-five degrees celsius.
Optionally, the temperature of the second electro-thermal film is set to sixty degrees celsius.
Optionally, the PCR amplification capillary comprises a silica gel capillary, a fluorinated ethylene propylene copolymer capillary, a polytetrafluoroethylene capillary, and a polyetheretherketone capillary.
Alternatively, the PCR amplification capillary is processed by an extrusion or thermal stretching process.
Optionally, the first electric heating film and the second electric heating film use metal foils and metal wires as inner electric conduction heating bodies, use a polyimide film as an outer insulation heat conduction layer, and the surface of the outer insulation heat conduction layer is provided with a back adhesive for fixing the PCR amplification capillary.
Optionally, the width ratio of the PCR amplification capillary wrapped by the first electrothermal film and the second electrothermal film is three to one.
Compared with the prior art, the invention adopting the technical scheme has the following technical effects: the invention relates to a portable liquid drop digital PCR device, which comprises a substrate, a movable sliding block, an oil phase injector, a water phase injector, a constant force spring, an injector fixing bracket, a connecting capillary tube, a liquid drop generating module, a PCR amplification capillary tube, a first electric heating film and a second electric heating film, the device comprises an injector fixing support, a movable sliding block, an oil-phase injector, a water-phase injector and a constant force spring, wherein the injector fixing support is fixed at one end of a substrate, the movable sliding block is arranged at the other end of the substrate, the injector fixing support is respectively connected with one ends of the oil-phase injector, the water-phase injector and the constant force spring, the movable sliding block is respectively connected with the other ends of the oil-phase injector, the water-phase injector and the constant force spring, one end of a connecting capillary tube is respectively connected with the oil-phase injector and the water-phase injector, the other end of the connecting capillary tube is connected with a liquid drop generation module, the output end of the liquid drop generation module is a PCR amplification capillary tube, and a first electric heating film and a second electric heating film wrap the PCR amplification capillary tube respectively. The injector is driven by the constant force spring, the system structure can be simplified, the liquid drops in the PCR amplification capillary are heated by the first electric heating film and the second electric heating film, the heating uniformity and stability of the liquid drops are improved, and the portability of the device can be realized.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.
FIG. 1 is a schematic structural diagram of a portable droplet digital PCR device according to an embodiment of the present invention;
fig. 2 is a schematic structural diagram of a droplet generation module of a portable droplet digital PCR device according to an embodiment of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present application more apparent, the present application is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the present application and are not intended to limit the present application.
Referring to fig. 1, an embodiment of the present invention provides a portable droplet digital PCR device, including a substrate 1, a movable slider 2, an oil-phase injector 3, a water-phase injector 4, a constant force spring 5, an injector fixing bracket 6, a connecting capillary 7, a droplet generation module 8, a first electrothermal film 11 and a second electrothermal film 12, wherein the droplet generation module 8 includes a PCR amplification capillary 9, the injector fixing bracket 6 is fixed at one end of the substrate 1, the movable slider 2 is disposed at the other end of the substrate 1, the injector fixing bracket 6 is respectively connected to the oil-phase injector 3, the water-phase injector 4 and one end of the constant force spring 5, the movable slider 2 is respectively connected to the oil-phase injector 3, the water-phase injector 4 and the other end of the constant force spring 5, one end of the connecting capillary 7 is respectively connected to the oil-phase injector 3 and the water-phase injector 4, the other end of the connecting capillary 7 is connected to the droplet generation module 8, the other end of the liquid drop generating module 8 is connected with a PCR amplification capillary 9, and the PCR amplification capillary 9 is respectively wrapped by a first electric heating film 11 and a second electric heating film 12.
The invention is arranged on a substrate through a movable sliding block and an injector fixing support, one end of a constant force spring is connected with the movable sliding block, the other end of the constant force spring is connected with the injector fixing support, an oil phase injector and a water phase injector are fixed on the injector fixing support, and tail pistons of the oil phase injector and the water phase injector are in contact with the movable sliding block. When the constant force spring is in a stretching state after the setting is finished, constant pulling force is provided for the movable sliding block, the movable sliding block drives the oil phase injector and the water phase injector, and the oil phase and the water phase in the injector enter the liquid drop generation module together through the connecting capillary. Wherein, the magnitude of the pulling force can be adjusted through the elastic coefficient of the constant force spring. Referring to fig. 2, the droplet generation module further includes a water phase capillary 15, an oil phase capillary 16, a PCR amplification capillary 9, and a sealant 17, and generates a micro droplet 13 through the droplet generation module, where the water phase capillary 15, the oil phase capillary 16, and the PCR amplification capillary 9 are all polymer capillaries, one end of the polymer capillary is an output end, the other end of the polymer capillary is an input end, an inner diameter of the output end is smaller than an inner diameter of the input end, the output end of the water phase capillary 15 is inserted into the output end of the PCR amplification capillary 9 from the input end of the PCR amplification capillary 9, the output end of the oil phase capillary 16 is connected to the input end of the PCR amplification capillary 9, and the sealant 17 is used to seal the input end of the PCR amplification capillary 9. The PCR amplification capillary 9 is an ultra-long pipeline for conveying micro-droplets, the input end of the PCR amplification capillary 99 is a pipeline with a thicker inner diameter, and the output end of the PCR amplification capillary 9 is a pipeline with a thinner inner diameter. The input ends of the water phase capillary 15 and the oil phase capillary 16 are connected with the connecting capillary, and the oil phase and the water phase in the oil phase injector and the water phase injector are driven to respectively enter the oil phase capillary 16 and the water phase capillary 15 by moving the sliding block. The output end of the oil phase capillary 16 is inserted into the input end of the PCR amplification capillary 9, and the oil phase enters the oil phase capillary 16, flows into the input end of the PCR amplification capillary 9 from the output end of the oil phase capillary 16, and flows to the output end of the PCR amplification capillary 9. The output end of the water phase capillary 15 is inserted into the output end of the PCR amplification capillary 9 from the input end of the PCR amplification capillary 9, and the water phase directly flows into the output end of the PCR amplification capillary 9 and forms water-in-oil droplets under the action of the cutting force of the oil phase with high peripheral flow velocity. The sealant is sealed at the input end of the PCR amplification capillary 9 and is used for preventing leakage and external air from entering.
Further as a preferred embodiment, referring to fig. 1, the device further includes a digital display temperature control console 10, the digital display temperature control console 10 is respectively connected to the first electric heating film 11 and the second electric heating film 12, and is used for controlling the temperature of the first electric heating film 11 and the second electric heating film 12.
The temperature of the first electric heating film is controlled to be 95 ℃, and the temperature of the second electric heating film is controlled to be 60 ℃. The real-time temperature of the first electric heating film and the second electric heating film can be observed through the digital display temperature control console, and the first electric heating film and the second electric heating film can be accurately controlled.
Further as a preferred embodiment, referring to fig. 1, the apparatus further includes a fluorescence detection module 14, the fluorescence detection module 14 includes a microscope camera, a filter and a computer, wherein the microscope camera is used for shooting and recording the state of the liquid drops flowing through the observation area, the filter is used for filtering light except for the fluorescent probe in the sample, and the computer is used for detecting the fluorescence intensity of each liquid drop passing through the observation area and counting the number of the liquid drops passing through the observation area.
The end of the output end of the PCR amplification capillary tube is an observation area, when the liquid drops are subjected to PCR amplification through 40 high-low temperature cycles, fluorescence intensity detection and liquid drop quantity statistics can be carried out on the liquid drops in the observation area through a fluorescence detection module, a microscope camera is used for shooting and recording the states of the liquid drops flowing through the observation area, an optical filter is further arranged in front of a lens of the microscope camera and used for filtering light except a fluorescence probe in a sample, the recorded data are uploaded to a computer through the microscope camera, the computer analyzes the uploaded data, the fluorescence intensity of each liquid drop passing through the observation area is detected, the number of the liquid drops passing through the observation area is counted, and finally, a chart is automatically drawn and the data are stored.
Further as a preferred embodiment, the temperature of the first electric heating film is set to ninety-five degrees celsius.
The temperature of the first electric heating film is set to ninety-five ℃, and the PCR amplification capillary tube area wrapped by the first electric heating film is a high-temperature area and used for enabling target DNA to be denatured in the high-temperature area and be uncoiled into two RNAs.
Further as a preferred embodiment, the temperature of the second electric heating film is set to sixty degrees celsius.
The temperature of the second electric heating film is set to sixty ℃, and the PCR amplification capillary tube area wrapped by the second electric heating film is a low-temperature area and is used for enabling RNA to be assembled into 2 new DNAs by using bases and fluorescent probes in a water phase, so that the amplification of the DNAs is completed.
Further as preferred embodiments, the PCR amplification capillary includes a silica gel capillary, a fluorinated ethylene propylene copolymer capillary, a polytetrafluoroethylene capillary, and a polyetheretherketone capillary.
The oil phase capillary, the water phase capillary and the PCR amplification capillary are all polymer capillaries, and the polymer capillaries comprise silica gel capillaries, fluorinated ethylene propylene copolymer capillaries, polytetrafluoroethylene capillaries and polyether ether ketone capillaries, have the advantage of good hydrophobicity, and can be better applied to generation of micro-droplets.
Further as a preferred embodiment, the PCR amplification capillary is processed by extrusion or thermal stretching process.
The oil-phase capillary, the water-phase capillary and the PCR amplification capillary are all polymer capillaries, and the polymer capillaries are obtained by extrusion or hot stretching process, so that the inner diameter and the outer diameter of the polymer capillaries can be better controlled, and the method is better applied to generation of micro-droplets.
Further as a preferred embodiment, the first electric heating film and the second electric heating film use metal foils and metal wires as inner electric conduction heating bodies, use polyimide films as outer insulating heat conduction layers, and the surfaces of the outer insulating heat conduction layers are provided with back glue for fixing PCR amplification capillary tubes.
After the liquid drops are generated by the liquid drop generation module, the liquid drops continuously flow in the PCR amplification capillary tube, sequentially pass through a high-temperature region (95 ℃) wrapped by the first electric heating film and a low-temperature region (60 ℃) wrapped by the second electric heating film, the first electric heating film and the second electric heating film use metal foils and metal wires as inner electric conduction heating bodies, use a polyimide film as an outer insulation heat conduction layer, the surface of the outer insulation heat conduction layer is provided with a back glue, the back glue is used for fixing the PCR amplification capillary tube, and the first electric heating film and the second electric heating film are folded after the PCR amplification capillary tube is fixed, so that the upper surface and the lower surface of the PCR amplification capillary tube are uniformly heated.
Further as a preferred embodiment, the width ratio of the first electric heating film and the second electric heating film to wrap the PCR amplification capillary is three to one.
Wherein, the time ratio required by the high-temperature area and the low-temperature area is 3: 1, the width ratio of the covered PCR amplification capillary in the first electric heating film and the second electric heating film is 3: 1, the time for the droplets to pass through the high temperature zone is 20 seconds, and the time for the droplets to pass through the low temperature zone is 60 seconds. The target DNA is denatured in the high temperature region and is deswirled into two RNAs, and the RNAs flow through the low temperature region and are assembled into 2 new DNAs by using bases in the water phase and a fluorescent probe, so that one round of amplification is completed. By analogy, after amplification is carried out for 40 cycles, the fluorescence intensity of the droplet containing the DNA is obviously enhanced, and finally the end of the PCR amplification capillary passes through a fluorescence detection region.
The invention relates to a portable liquid drop digital PCR device, which comprises the following steps: the water phase injector and the oil phase injector are driven by the movable sliding block and the constant force spring, the magnitude of the pulling force is controlled by the constant force spring, so that the water phase and the oil phase in the water phase injector and the oil phase injector enter the liquid drop generating module through the connecting capillary, and the water phase and the oil phase generate liquid drops through the liquid drop generating module and enter the PCR amplification capillary. The temperature of the first electric heating film and the temperature of the second electric heating film are controlled through a digital display temperature control console, a high-temperature area and a low-temperature area are formed in the PCR amplification capillary, the liquid drop passes through the high-temperature area and the low-temperature area to carry out PCR amplification reaction, and finally, the end of the PCR amplification capillary passes through a fluorescence detection area, namely an observation area, and the fluorescence detection is carried out on the liquid drop through a fluorescence detection module.
In the related technology, the injection module mostly adopts a mechanical pump or a pneumatic pump to drive the water phase and the oil phase, so that the injection module has larger volume and high cost; one side of the amplification pipeline is heated by using a heating table, a heat conduction module or a heating sheet in the PCR amplification module, so that the temperature of the side close to a heat source is higher than that of the other side, and the pipeline is heated unevenly. In addition, both of the single heat source and the multi-heat source systems require a heating table or a heating plate having a large volume, and thus are not portable and lightweight.
In summary, the embodiments of the present invention have the following advantages:
(1) the embodiment of the invention drives the injector through the constant force spring, simplifies the system structure, reduces the volume and the weight of the system, reduces the equipment cost and realizes the portability of the device;
(2) according to the embodiment of the invention, the folded electrothermal film is used for carrying out wrapped heating on micro liquid drops flowing in the capillary tube, so that the uniformity and the stability of a heated area are improved, the problem of influence of environmental temperature is solved, the heated areas of the high-temperature area and the low-temperature area are controlled, the heated time ratios of different temperature areas are accurately controlled, and the target DNA is ensured to be amplified smoothly;
(3) compared with a heating table, the electric heating film in the embodiment of the invention greatly reduces the volume and the weight of the system, and further realizes the portability of the device.
In alternative embodiments, the functions/acts noted in the block diagrams may occur out of the order noted in the operational illustrations. For example, two blocks shown in succession may, in fact, be executed substantially concurrently, or the blocks may sometimes be executed in the reverse order, depending upon the functionality/acts involved. Furthermore, the embodiments presented and described in the flow charts of the present invention are provided by way of example in order to provide a more thorough understanding of the technology. The disclosed methods are not limited to the operations and logic flows presented herein. Alternative embodiments are contemplated in which the order of various operations is changed and in which sub-operations described as part of larger operations are performed independently.
Furthermore, although the present invention is described in the context of functional modules, it should be understood that, unless otherwise stated to the contrary, one or more of the functions and/or features may be integrated in a single physical device and/or software module, or one or more functions and/or features may be implemented in separate physical devices or software modules. It will also be appreciated that a detailed discussion of the actual implementation of each module is not necessary for an understanding of the present invention. Rather, the actual implementation of the various functional modules in the apparatus disclosed herein will be understood within the ordinary skill of an engineer, given the nature, function, and internal relationship of the modules. Accordingly, those skilled in the art can, using ordinary skill, practice the invention as set forth in the claims without undue experimentation. It is also to be understood that the specific concepts disclosed are merely illustrative of and not intended to limit the scope of the invention, which is defined by the appended claims and their full scope of equivalents.
The functions, if implemented in the form of software functional units and sold or used as a stand-alone product, may be stored in a computer readable storage medium. Based on such understanding, the technical solution of the present invention may be embodied in the form of a software product, which is stored in a storage medium and includes instructions for causing a computer device (which may be a personal computer, a server, or a network device) to execute all or part of the steps of the method according to the embodiments of the present invention. And the aforementioned storage medium includes: a U disk, a removable hard disk, a Read-only Memory (ROM), a Random Access Memory (RAM), a magnetic disk or an optical disk, and other various media capable of storing program codes.
The logic and/or steps represented in the flowcharts or otherwise described herein, e.g., an ordered listing of executable instructions that can be considered to implement logical functions, can be embodied in any computer-readable medium for use by or in connection with an instruction execution system, apparatus, or device, such as a computer-based system, processor-containing system, or other system that can fetch the instructions from the instruction execution system, apparatus, or device and execute the instructions. For the purposes of this description, a "computer-readable medium" can be any means that can contain, store, communicate, propagate, or transport the program for use by or in connection with the instruction execution system, apparatus, or device.
More specific examples (a non-exhaustive list) of the computer-readable medium would include the following: an electrical connection (electronic device) having one or more wires, a portable computer diskette (magnetic device), a Random Access Memory (RAM), a read-only memory (ROM), an erasable programmable read-only memory (EPROM or flash memory), an optical fiber device, and a portable compact disc read-only memory (CDROM). Additionally, the computer-readable medium could even be paper or another suitable medium upon which the program is printed, as the program can be electronically captured, via for instance optical scanning of the paper or other medium, then compiled, interpreted or otherwise processed in a suitable manner if necessary, and then stored in a computer memory.
It should be understood that portions of the present invention may be implemented in hardware, software, firmware, or a combination thereof. In the above embodiments, the various steps or methods may be implemented in software or firmware stored in memory and executed by a suitable instruction execution system. For example, if implemented in hardware, as in another embodiment, any one or combination of the following techniques, which are known in the art, may be used: a discrete logic circuit having a logic gate circuit for implementing a logic function on a data signal, an application specific integrated circuit having an appropriate combinational logic gate circuit, a Programmable Gate Array (PGA), a Field Programmable Gate Array (FPGA), or the like.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
While embodiments of the invention have been shown and described, it will be understood by those of ordinary skill in the art that: various changes, modifications, substitutions and alterations can be made to the embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents.
While the preferred embodiments of the present invention have been illustrated and described, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (9)
1. The utility model provides a portable liquid drop digit PCR device, its characterized in that, includes basement (1), removes slider (2), oil phase syringe (3), aqueous phase syringe (4), constant force spring (5), syringe fixed bolster (6), connects capillary (7), liquid drop generation module (8), first electric heat membrane (11) and second electric heat membrane (12), wherein, liquid drop generation module (8) are including PCR amplifys capillary (9), syringe fixed bolster (6) are fixed the one end of basement (1), it arranges to remove slider (2) the other end of basement (1), syringe fixed bolster (6) are connected respectively oil phase syringe (3), aqueous phase syringe (4) and the one end of constant force spring (5), it connects respectively to remove slider (2) oil phase syringe (3), The water phase injector (4) with the other end of constant force spring (5), connect capillary (7) one end and connect respectively oil phase injector (3) with water phase injector (4), connect capillary (7) other end and connect liquid drop generation module (8), the output of liquid drop generation module (8) is PCR amplification capillary (9), first electric heat membrane (11) with second electric heat membrane (12) is wrapping up respectively PCR amplification capillary (9).
2. The portable liquid drop digital PCR device according to claim 1, further comprising a digital display temperature control console (10), wherein the digital display temperature control console (10) is respectively connected to the first electrothermal film (11) and the second electrothermal film (12) for controlling the temperature of the first electrothermal film (11) and the second electrothermal film (12).
3. The portable droplet digital PCR device according to claim 1, further comprising a fluorescence detection module (14), wherein the fluorescence detection module (14) comprises a microscope camera, a filter and a computer, wherein the microscope camera is used for shooting and recording the state of the droplets flowing through the observation area, the filter is used for filtering light in the sample except the fluorescent probe, and the computer is used for detecting the fluorescence intensity of each droplet passing through the observation area and counting the number of the droplets passing through the observation area.
4. A portable droplet digital PCR device according to claim 1, wherein the temperature of the first electro-thermal film (11) is set to ninety-five degrees celsius.
5. A portable droplet digital PCR device according to claim 1, wherein the temperature of the second electro-thermal film (12) is set to sixty degrees celsius.
6. The portable droplet digital PCR device according to claim 4, wherein the PCR amplification capillary (9) comprises a silica gel capillary, a fluorinated ethylene propylene copolymer capillary, a polytetrafluoroethylene capillary and a polyetheretherketone capillary.
7. The portable droplet digital PCR device according to claim 6, wherein the PCR amplification capillary (9) is processed by extrusion or hot-drawing process.
8. The portable liquid drop digital PCR device according to claim 1, wherein the first electric heating film (11) and the second electric heating film (12) use metal foil and metal wire as inner electric heating conductive bodies, use polyimide film as outer insulating heat conducting layer, and the surface of the outer insulating heat conducting layer is provided with a back adhesive for fixing the PCR amplification capillary (9).
9. The portable droplet digital PCR device according to claim 1, wherein the width ratio of the first electric heating film (11) and the second electric heating film (12) to wrap the PCR amplification capillary (9) is three to one.
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