Integrated digital PCR machine

Abstract

Provided is an integrated digital PCR machine, comprising: a temperature control module, which can control the temperature of a reaction zone of a microfluidic chip located at a bearing position thereof, so as to perform a PCR reaction; an optical detection module, which can detect a PCR reaction result in the reaction zone of the microfluidic chip; a pressure-plate module, which comprises an air supply assembly, wherein when moving above the temperature control module, the air supply assembly can output air pressure into an inlet of the microfluidic chip, which is located at the bearing position of the temperature control module, or can stop outputting air pressure; and a chip moving module, which can move the microfluidic chip from the bearing position of the temperature control module to a detection position of the optical detection module. By means of the provided integrated digital PCR machine, an instrument can automatically produce droplets and perform PCR reactions and optical scanning detection by means of simply adding a specimen into a microfluidic chip and loading the chip into the integrated digital PCR machine, thereby greatly improving the convenience of use of the instrument.

Description

Digital PCR all-in-one machine TECHNICAL FIELD

[0001] The present application relates to the technical field of high-throughput analysis systems, and in particular to a digital PCR all-in-one machine. BACKGROUND

[0002] Microfluidics is a technology for precisely controlling and manipulating microscale fluid, which has the characteristics of small capacity (nanoliter, picoliter, and femtoliter level) and low energy consumption. People use microfluidic technology to integrate the basic operation units of biological, chemical, and medical analysis processes, such as sample preparation, reaction, separation, and detection, onto a microfluidic chip with microfluidic channels as the basic structure, thereby achieving rapid sample processing and detection, and reducing the use of reagents and samples. Due to its great potential in the fields of biology, chemistry, medicine, and others, microfluidics has become a new research field that combines biology, chemistry, medicine, fluid, electronics, materials, and mechanics.

[0003] Digital PCR, a nucleic acid quantification method based on single-molecule PCR counting, is an absolute quantification method. The principle is to disperse a large amount of diluted nucleic acid solution into microreactors or microdroplets on a microfluidic chip by microfluidic or microdropletization, so that the number of nucleic acid templates in each microreactor is less than or equal to one. After PCR cycles, a reactor or microdroplet with one nucleic acid molecule template will give a fluorescence signal, and a reactor or microdroplet without a template will have no fluorescence signal. According to the relative proportion and the volume of the reactor, the nucleic acid concentration of the original solution can be calculated.

[0004] In the existing digital PCR system, the user moves the generated droplets to the PCR reaction test tube with a pipette, then puts the test tube into the PCR thermal cycler for PCR reaction, and after the reaction is completed, the user takes out the PCR tube and puts it into the droplet fluorescence detector for droplet fluorescence detection. The detection principle is to put the droplets and oil into a capillary, and the droplets pass through the detection position one by one, and the instrument reads whether the droplets have fluorescence. However, this system has the following disadvantages: (1) the user needs to manually transfer the droplets to the 96-well plate after the system generates the droplets, which can cause loss or rupture of the droplets; (2) due to the limitation of heat transfer rate (the sample in the test tube is thick, and heating starts from the outside of the test tube), the PCR reaction in the test tube is slow, so the entire PCR process takes a long time, usually 150 minutes; (3) after the droplets pass through the detection position, they enter the waste liquid bottle, and the sample cannot be detected again.

[0005] Currently, there is no automatic PCR instrument specially designed for microfluidic chips on the market. SUMMARY

[0006] To solve the problems in the prior art, the application provides a digital PCR all-in-one machine. The technical scheme of the application is as follows:

[0007] 1. A digital PCR all-in-one machine, comprising:

[0008] a temperature control module, a bearing position of the temperature control module being capable of bearing a microfluidic chip or a tray for loading the microfluidic chip, the temperature control module being capable of controlling the temperature of a reaction zone of the microfluidic chip located at the bearing position of the temperature control module to perform a PCR reaction;

[0009] an optical detection module, the optical detection module being arranged on one side of the temperature control module and being capable of detecting a PCR reaction result in the reaction zone of the microfluidic chip;

[0010] a pressing plate module, the pressing plate module comprising a gas supply assembly, the gas supply assembly being capable of moving to and moving out of the upper part of the temperature control module; when the gas supply assembly moves to the upper part of the temperature control module, the gas supply assembly is capable of outputting or stopping outputting air pressure to the inlet of the microfluidic chip located at the bearing position of the temperature control module;

[0011] a chip moving module, the chip moving module being capable of moving the microfluidic chip from the bearing position of the temperature control module to a detection position of the optical detection module, so that the optical detection module can detect the microfluidic chip.

[0012] 2. The digital PCR all-in-one machine of item 1, wherein,

[0013] the temperature control module comprises a first temperature control assembly, the first temperature control assembly being capable of controlling the temperature of at least one surface of the microfluidic chip located at the bearing position of the temperature control module.

[0014] 3. The digital PCR all-in-one machine of item 2, wherein,

[0015] the first temperature control assembly comprises one or more first temperature control units, each of the first temperature control units corresponding to a reaction zone of the microfluidic chip.

[0016] 4. The digital PCR all-in-one machine of item 2, wherein,

[0017] the temperature control module further comprises a first heat dissipation assembly;

[0018] the first heat dissipation assembly comprises:

[0019] a first heat dissipation channel, the first heat dissipation channel being located at the bottom of the first temperature control assembly; and,

[0020] a heat dissipation fan, the heat dissipation fan being arranged in the first heat dissipation channel.

[0021] 5. The digital PCR all-in-one machine according to item 1, wherein,

[0022] the optical detection module comprises:

[0023] an optical detection assembly capable of detecting the PCR reaction result in the reaction zone of the microfluidic chip;

[0024] an optical detection moving assembly capable of moving the optical detection assembly to detect the PCR reaction result in different microfluidic chips and / or different reaction zones of the microfluidic chip.

[0025] 6. The digital PCR all-in-one machine according to item 5, wherein,

[0026] the optical detection assembly comprises:

[0027] an excitation light emitting unit capable of emitting excitation light to the microfluidic chip moved to the detection position;

[0028] a light signal receiving unit capable of receiving fluorescence from the microfluidic chip and converting the fluorescence into an electrical signal.

[0029] 7. The digital PCR all-in-one machine according to item 6, wherein,

[0030] the excitation light emitting unit comprises:

[0031] an excitation light emitting subunit capable of emitting excitation light;

[0032] a first light filtering subunit provided with two or more first light filters, each of the first light filters capable of filtering the excitation light emitted by the excitation light emitting subunit to obtain excitation light of different wave bands, respectively;

[0033] a first switching subunit capable of moving and / or rotating the first light filtering subunit to switch different first light filters to the optical path of the excitation light emitting subunit.

[0034] 8. The digital PCR all-in-one machine according to item 7, wherein,

[0035] the excitation light emitting subunit comprises an excitation light source and a first light beam shaping lens group arranged in the optical path of the excitation light source.

[0036] 9. The digital PCR all-in-one machine according to item 6, wherein,

[0037] the light signal receiving unit comprises:

[0038] a light signal receiving subunit, which is capable of converting fluorescence from the microfluidic chip into an electrical signal;

[0039] a second light filtering subunit, which is provided with two or more second light filters, and each of the second light filters is capable of filtering fluorescence from the microfluidic chip to obtain fluorescence of different wave bands, respectively;

[0040] a second switching subunit, which is capable of moving and / or rotating the second light filtering subunit to switch different second light filters to the receiving light path of the light signal receiving subunit.

[0041] 10. The digital PCR all-in-one machine according to claim 9, wherein,

[0042] the light signal receiving unit further comprises:

[0043] a light receiving subunit, which is arranged between the second light filtering subunit and the microfluidic chip, and the light receiving subunit comprises a light shielding cylinder and a second light beam shaping lens group arranged in the light shielding cylinder.

[0044] 11. The digital PCR all-in-one machine according to claim 7, wherein,

[0045] the optical detection assembly further comprises:

[0046] a complementary color unit, which comprises a plurality of complementary color light emitting subunits, the complementary color light exciting unit is capable of emitting monochromatic light, and the wave bands of the monochromatic light emitted by different complementary color light emitting subunits and the excitation light filtered by different first light filtering subunits are all different.

[0047] 12. The digital PCR all-in-one machine according to claim 11, wherein,

[0048] the complementary color light emitting subunit comprises a complementary color light source, a third light beam shaping lens group arranged on the light path of the complementary color light source, and a third light filter.

[0049] 13. The digital PCR all-in-one machine according to claim 1, wherein,

[0050] when the gas supply assembly is moved to the upper part of the temperature control module, the gas supply assembly is further capable of outputting or stopping outputting air pressure to the outlet of the microfluidic chip located in the carrying position of the temperature control module.

[0051] 14. The digital PCR all-in-one machine according to claim 1, wherein,

[0052] the pressing plate module further comprises:

[0053] The first pressing plate moving assembly is capable of moving the gas supply assembly to and from the upper part of the temperature control module.

[0054] 15. The digital PCR all-in-one machine according to item 14, wherein,

[0055] The first pressing plate moving assembly comprises:

[0056] A bearing frame capable of bearing the gas supply assembly;

[0057] A first guide rail arranged on the rack;

[0058] A sliding block arranged at the bottom of the bearing frame;

[0059] A first motion unit, the moving end of which is directly or indirectly connected with the sliding block, so as to drive the gas supply assembly arranged on the bearing frame to slide along the first guide rail through the sliding block.

[0060] 16. The digital PCR all-in-one machine according to item 15, wherein,

[0061] The pressing plate module further comprises:

[0062] A second pressing plate moving assembly, which is capable of moving the gas supply assembly to the gas supply position of the microfluidic chip or moving the gas supply assembly away from the gas supply position of the microfluidic chip when the first pressing plate moving assembly moves the gas supply assembly to the upper part of the temperature control module.

[0063] 17. The digital PCR all-in-one machine according to item 16, wherein,

[0064] The second pressing plate moving assembly comprises:

[0065] A second motion unit, the moving end of which is directly or indirectly connected with the gas supply assembly, so as to drive the gas supply assembly to move to the gas supply position of the microfluidic chip or move away from the gas supply position of the microfluidic chip.

[0066] 18. The digital PCR all-in-one machine according to item 17, wherein,

[0067] The bearing frame comprises:

[0068] A fixed plate; and

[0069] A guide column connecting the sliding block and the fixed plate;

[0070] The second pressing plate moving assembly further comprises:

[0071] A movable plate is provided with a guide hole, the guide hole is sleeved on the guide column, the movable plate can slide along the guide column; the fixed end of the second movement unit is connected with the fixed plate; the movement end of the second movement unit is connected with the movable plate; the air supply assembly is fixed at the bottom of the movable plate.

[0072] 19. The digital PCR all-in-one machine of item 17, wherein,

[0073] The pressing plate module further comprises:

[0074] A second heating assembly, when the second pressing plate moving assembly drives the air supply assembly to move to the air supply position of the microfluidic chip, the second heating assembly can heat the opposite side of the temperature control module of the microfluidic chip.

[0075] 20. The digital PCR all-in-one machine of item 19, wherein,

[0076] The upper end of the second heating assembly is connected with an elastic member, and the elastic member is directly or indirectly fixedly connected with the movement end of the second movement unit.

[0077] The air outlet of the air supply assembly is located on both sides of the second heating assembly.

[0078] 21. The digital PCR all-in-one machine of item 19, wherein,

[0079] The second heating assembly comprises one or more second heating units.

[0080] Each second heating unit corresponds to a reaction area of the microfluidic chip.

[0081] 22. The digital PCR all-in-one machine of item 21, wherein,

[0082] The second heating unit is provided with a plurality of blind holes facing the reaction area of the microfluidic chip.

[0083] 23. The digital PCR all-in-one machine of item 1, wherein,

[0084] The pressing plate module is provided with two oppositely arranged limiting grooves;

[0085] The pressing plate module further comprises a baffle, and the baffle comprises:

[0086] A baffle body, the baffle body is located on the inner side of the oppositely arranged limiting grooves;

[0087] Two or more first protrusions, two or more first protrusions are arranged on both sides of the baffle body, and the first protrusions are respectively located in the limiting grooves;

[0088] two or more second protrusions, the two or more second protrusions are arranged on the bottom of the baffle body;

[0089] wherein, when the chip moving module moves the microfluidic chip to the detection position of the optical detection module, the baffle separates the microfluidic chip.

[0090] 24. The digital PCR all-in-one machine of item 18, wherein,

[0091] The pressing plate module further comprises:

[0092] a resilient component, which is directly or indirectly fixed to the bottom of the movable plate, and when the gas supply component is at the gas supply position of the microfluidic chip, the bottom of the resilient component is at the top of the microfluidic chip or a tray for loading the microfluidic chip and is in a stressed state.

[0093] 25. The digital PCR all-in-one machine of item 16, wherein,

[0094] The gas supply component is capable of moving between the loading position of the temperature control module and the detection position of the optical detection module.

[0095] The chip moving module is directly or indirectly fixedly connected with the gas supply component.

[0096] 26. The digital PCR all-in-one machine of item 25, wherein,

[0097] The chip moving module comprises:

[0098] a chip carrier assembly, one end of the chip carrier assembly is directly or indirectly fixedly connected with the gas supply component, and the other end extends to the lower part of the gas supply component, so that when the second pressing plate moving assembly moves the gas supply component out of the gas supply position of the microfluidic chip, the chip carrier assembly can lift a tray for loading the microfluidic chip, so that the microfluidic chip leaves the loading position of the temperature control module.

[0099] 27. The digital PCR all-in-one machine of item 26, wherein,

[0100] The chip carrier assembly at least comprises:

[0101] a pair of chip carrier bodies, one end of the chip carrier body is directly or indirectly fixedly connected with the gas supply component;

[0102] a third movement unit, which is arranged on the chip carrier body;

[0103] A clamping jaw is arranged at the moving end of the third moving unit, and when the gas supply assembly is located at the gas supply position of the microfluidic chip, the moving end of the third moving unit can drive the clamping jaw to clamp the tray for loading the microfluidic chip.

[0104] 28. The digital PCR all-in-one machine of item 26, wherein,

[0105] The chip tray assembly comprises:

[0106] Two chip tray units are arranged in pairs, one end of the chip tray unit is directly or indirectly fixedly connected with the gas supply assembly, and the other end of the chip tray unit extends to the lower part of the gas supply assembly.

[0107] 29. The digital PCR all-in-one machine of item 28, wherein,

[0108] The chip tray assembly further comprises:

[0109] A third protrusion is arranged on the upper surface of the other end of the chip tray unit.

[0110] The above-mentioned digital PCR all-in-one machine provided by the present application only needs to add samples into the microfluidic chip and load the chip into the digital PCR all-in-one machine, and the instrument can automatically complete droplet generation, PCR reaction and optical scanning detection, greatly increasing the convenience of instrument use.

[0111] The above description is only a summary of the technical solutions of the present application. In order to make the technical means of the present application more clear and understandable, and to the extent that the contents of the specification can be implemented by those skilled in the art, and in order to make the above and other purposes, features and advantages of the present application more obvious and easy to understand, the following will be illustrated with specific embodiments. BRIEF DESCRIPTION OF DRAWINGS

[0112] Figure 1: a side view structural schematic diagram of the digital PCR all-in-one machine in one embodiment of the present application;

[0113] Figure 2: a side view structural schematic diagram of the digital PCR all-in-one machine in one embodiment of the present application;

[0114] Figure 3: a side view structural schematic diagram of the digital PCR all-in-one machine in one embodiment of the present application;

[0115] Figure 4: a structural schematic diagram of the digital PCR all-in-one machine in one embodiment of the present application;

[0116] Figure 5: a structural schematic diagram of the chip moving module arrangement in the digital PCR all-in-one machine in one embodiment of the present application;

[0117] Figure 6: a schematic diagram of the chip moving module structure in the digital PCR all-in-one machine according to an embodiment of the present application;

[0118] Figure 7: a schematic diagram of the chip moving module setting mode structure in the digital PCR all-in-one machine according to another embodiment of the present application;

[0119] Figure 8: a schematic diagram of the chip moving module structure in the digital PCR all-in-one machine according to another embodiment of the present application;

[0120] Figure 9: a schematic diagram of the air supply assembly located at the air supply position of the microfluidic chip in the digital PCR all-in-one machine according to an embodiment of the present application;

[0121] Figure 10: a schematic diagram of the second temperature control unit in the digital PCR all-in-one machine according to an embodiment of the present application;

[0122] Figure 11: a bottom view of the second temperature control unit in the digital PCR all-in-one machine according to an embodiment of the present application;

[0123] Figure 12: a schematic diagram of the air supply assembly leaving the air supply position of the microfluidic chip in the digital PCR all-in-one machine according to an embodiment of the present application;

[0124] Figure 13: a schematic diagram of the baffle position when the air supply assembly is located at the air supply position of the microfluidic chip according to an embodiment of the present application;

[0125] Figure 14: a schematic diagram of the baffle position when the air supply assembly is located at the detection position of the optical detection module according to an embodiment of the present application;

[0126] Figure 15: a schematic diagram of the microfluidic chip and the first temperature control unit in the digital PCR all-in-one machine according to an embodiment of the present application;

[0127] Figure 16: a schematic diagram of the optical detection assembly according to an embodiment of the present application;

[0128] Figure 17: a schematic diagram of part of the structure of Figure 16;

[0129] Figure 18: a schematic diagram of part of the structure of Figure 16;

[0130] Figure 19: a schematic diagram of the optical detection assembly according to another embodiment of the present application;

[0131] Figure 20: a schematic diagram of part of the structure of Figure 19;

[0132] Figure 21: a schematic diagram of part of the structure of Figure 19;

[0133] Explanation of reference signs: 100, temperature control module; 110, first temperature control assembly; 111, first temperature control unit; 120, first heat dissipation assembly; 200, optical detection module; 210, optical detection assembly; 210-110, excitation light emitting subunit; 210-111, excitation light source; 210-112, first beam shaping lens group; 210-120, first light filtering subunit; 210-121, first light filtering wheel; 210-122, first light filter; 210-130, first switching subunit; 210-140, first heat dissipation member; 210-220, second light filtering subunit; 210-221, second light filtering wheel; 210-222, second light filter; 210-230, light receiving subunit; 210-231, light shielding cylinder; 210-232, second beam shaping lens group; 210-310, complementary color light emitting subunit; 210-311, complementary color light source; 210-312, third beam shaping lens group; 210-313, third light filter; 210-410, first fixed plate; 210-420, second fixed plate; 210-500, second heat dissipation member; 220, optical detection moving assembly; 221, first motor; 222, first screw rod; 230, second heat dissipation channel; 300, pressing plate module; 310, air supply assembly; 321, bearing frame; 322, first guide rail; 323, sliding block; 324, first movement unit; 325, second motor; 326, second screw rod; 327, fixed plate; 328, guide column; 331, second movement unit; 332, movable plate; 333, third motor; 334, third screw rod; 340, second heating assembly; 341, elastic member; 342, second heating unit; 343, blind hole; 350, limiting groove; 360, baffle; 361, baffle body; 362, first protrusion; 363, second protrusion; 370, elastic assembly; 400, chip moving module; 410, chip carrier body; 420, third movement unit; 430, clamping jaw; 440, chip carrier unit; 450, third protrusion; 500, microfluidic chip; 600, tray; 700, control module; 800, display; 900, rack. DETAILED DESCRIPTION

[0134] The following embodiments of the present application are only used to illustrate the specific embodiments of the present application, and these embodiments cannot be understood as limiting the present application. Any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application are equivalent replacement methods, which fall within the protection scope of the present application.

[0135] It should be understood by those skilled in the art that in the disclosure of the present application, the terms "first", "second", "third", "fourth", "fifth" and the like are only used to distinguish different structures, without limiting the number, connection relationship, etc. of specific structures; in addition, the orientation or positional relationship indicated by "longitudinal", "transverse", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer" and the like is based on the orientation or positional relationship shown in the drawings, which is only for the convenience of describing the present application and simplifying the description, and does not indicate or imply that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, therefore the above-mentioned terms cannot be understood as a limitation on the present application.

[0136] The present embodiment provides a digital PCR all-in-one machine, as shown in FIGS. 1-15, comprising:

[0137] A temperature control module 100, the carrying position (i.e. the position of carrying the microfluidic chip 500 or the tray 600) of the temperature control module 100 can carry the microfluidic chip 500 or carry the tray 600 for loading the microfluidic chip (specifically, in the present embodiment, the tray 600 is carried, and the microfluidic chip 500 is loaded on the tray 600), and the temperature control module 100 can control the temperature of the reaction area (the area where the reaction cavity is arranged) of the microfluidic chip 500 located in the carrying position of the temperature control module 100 to perform PCR reaction;

[0138] An optical detection module 200, the optical detection module 200 can detect the PCR reaction result in the reaction area of the microfluidic chip 500;

[0139] A pressing plate module 300, the pressing plate module 300 comprises a gas supply assembly 310, the gas supply assembly 310 can move to and move out of the upper part of the temperature control module 100 (in the present embodiment, specifically, it moves in the horizontal direction); when the gas supply assembly 310 moves to the upper part of the temperature control module 100, the gas supply assembly 310 can output or stop outputting air pressure to the inlet of the microfluidic chip 500 located in the carrying position of the temperature control module 100, so as to make the oil phase and the water phase in the microfluidic chip 500 move from the inlet to the outlet direction of the microfluidic chip 500, to form droplets in the reaction cavity of the microfluidic chip 500 for performing PCR;

[0140] A chip moving module 400, the chip moving module 400 can move the microfluidic chip 500 from the carrying position of the temperature control module 100 to the detection position (i.e. the position where the optical detection module 200 can detect the microfluidic chip 500) of the optical detection module 200 (in the present embodiment, specifically, it moves in the vertical direction), so as to facilitate the optical detection module 200 to detect the microfluidic chip 500.

[0141] Further, in the embodiment, when the pressing plate module 300 moves to the upper part of the temperature control module 100, the gas supply assembly 310 can also output gas pressure or stop outputting gas pressure to the outlet of the microfluidic chip 500 located in the carrying position of the temperature control module 100. Thus, when the chip moving module 400 moves the microfluidic chip 500 from the temperature control module 100 to the detection position of the optical detection module 200, the inlet and outlet of the microfluidic chip 500 can be pressurized at the same time to prevent the movement of droplets or the evaporation of liquid (oil phase and water phase), so as to improve the accuracy of detection.

[0142] Specifically, in the embodiment, the gas supply assembly 310 is a gas supply pressing plate, the inside of the gas supply pressing plate is provided with a gas flow channel, and the lower surface of the gas supply pressing plate is provided with gas outlets corresponding to the inlet and outlet of the microfluidic chip, and the gas supply pressing plate is provided with a gas inlet hole connected with a gas source to provide gas with different pressures, so that the pneumatic pressing plate can provide different pressures for the oil phase and the water phase. Generally, the generation of droplets adopts a two-step method: in the first step, the oil phase pressure is much greater than the water phase pressure, so that the oil pre-fills the flow channel / chamber, stabilizes the surface properties of the flow channel wall during the droplet generation process, and thus benefits the generation of droplets with consistent size and stability; in the second step, specific pressures are set for the oil phase and the water phase according to the size of the required droplets, so as to generate droplets with specific size or adjust the size of the droplets. The gas supply pressing plate can provide different pressures for the inlet (oil phase and water phase) and outlet of the chip, so as to control the movement of droplets, adjust the relative position of the droplets in the chip chamber and the density of the droplets; or the gas supply pressing plate can also provide the same pressure, so as to increase the boiling point of water and the solubility of gas in liquid, thereby preventing the generation of bubbles, preventing the evaporation of liquid (water phase or oil phase) in the chamber and the movement of droplets.

[0143] In use of the digital PCR all-in-one machine provided in the embodiment, first, the pressing plate module 300 is not in the upper part of the temperature control module 100, and in the embodiment, the pressing plate module 300 is in the upper part of the optical detection module 200 (as shown in FIG. 1, FIG. 3), so as to expose the temperature control module 100, and the microfluidic chip 500 loaded with liquid phase and oil phase is carried to the carrying position of the temperature control module 100 through the tray 600; then, the pressing plate module 300 is moved to the upper part of the temperature control module 100, and the gas supply assembly 310 outputs gas pressure to the inlet of the microfluidic chip 500 located in the carrying position of the temperature control module 100, until the oil phase and the water phase in the microfluidic chip 500 move from the inlet to the outlet of the microfluidic chip 500, and droplets for performing PCR are formed in the reaction chamber of the microfluidic chip 500; the temperature control module 100 controls the temperature of the reaction area of the microfluidic chip 500 (heating / cooling) to perform PCR reaction; then, the chip moving module 400 moves the microfluidic chip 500 from the temperature control module 100 to the detection position of the optical detection module 200, and the optical detection module 200 is used to detect the microfluidic chip 500.

[0144] By using the digital PCR all-in-one machine of the present application, only the sample needs to be added to the microfluidic chip and the chip is loaded into the digital PCR all-in-one machine, and the instrument can automatically complete droplet generation, PCR reaction and optical scanning detection, greatly increasing the convenience of instrument use.

[0145] In one embodiment, as shown in FIG. 1, FIG. 15, the temperature control module 100 comprises a first temperature control assembly 110 for temperature control of at least one surface of the microfluidic chip 500 located in the temperature control module 100 carrying position (in this embodiment, the lower surface of the microfluidic chip 500).

[0146] Specifically, in this embodiment, the first temperature control assembly 110 comprises one or more (such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20 or more, in this embodiment, 8) first temperature control units 111, each of which corresponds to a reaction zone of the microfluidic chip 500, thereby enabling independent temperature control of each microfluidic chip 500. Independent first temperature control units 111 can allow different temperatures to be set, such as forming a temperature gradient on chips tested in parallel, enabling a screening of suitable temperatures required for PCR reaction, or screening and research of other temperature-sensitive conditions.

[0147] Regarding the first temperature control unit 111, the present application is not specifically limited as long as it can heat the microfluidic chip 500. Preferably, the first temperature control unit 111 can not only heat the microfluidic chip 500, but also cool the microfluidic chip 500, thereby enabling faster and more accurate temperature control to facilitate accurate PCR reaction under specific temperature conditions. In this embodiment, the first temperature control unit 111 is a TEC heating unit (semiconductor cooler, also known as thermoelectric cooler) to enable heating and cooling of the microfluidic chip 500.

[0148] Further, in this embodiment, the temperature control module 100 further comprises a first heat dissipation assembly 120, which comprises a first heat dissipation channel at the bottom of the first temperature control assembly 110 and a heat dissipation fan (not shown in the figure) arranged in the first heat dissipation channel. Thus, when cooling is needed, the first heat dissipation assembly 120 can more quickly remove the heat of the first temperature control assembly 110 and the microfluidic chip 500, facilitating rapid cooling, thereby facilitating more accurate and rapid PCR reaction conditions and accurate PCR reaction under specific temperature conditions.

[0149] In one embodiment, as shown in FIG. 1-3, the optical detection module 200 comprises:

[0150] an optical detection assembly 210 capable of detecting the PCR reaction result in the reaction area of the microfluidic chip 500;

[0151] an optical detection moving assembly 220 capable of moving the optical detection assembly 210 to detect the PCR reaction result in different microfluidic chips 500 and / or different reaction chambers of a microfluidic chip 500.

[0152] Regarding the optical detection assembly, it can specifically be a lens unit, a filter, and a photosensitive chip (CMOS sensor) arranged in sequence from one side to the other side to take a photo of the reaction area of the microfluidic chip (e.g., take a photo of the fluorescence after PCR reaction), so as to obtain the detection result according to the photo, including the size of the droplet and the fluorescence intensity of the droplet, etc.

[0153] Regarding the optical detection moving assembly 220, the present application does not have specific limitations as long as it can move the optical detection assembly 210. As shown in FIG. 3, in the present embodiment, the optical detection assembly is moved by a lead screw, specifically, a motor (first motor 221) drives the rotation of the screw rod (first screw rod 222) of the lead screw through a belt, so as to drive the nut arranged on the screw rod to move, so as to drive the optical detection assembly 210 fixedly arranged with the nut to move. More specifically, in the present embodiment, the arrangement direction of the screw rod (first screw rod 222) of the lead screw is perpendicular to the direction in which the chip moving module drives the microfluidic chip to move, so as to be capable of detecting each microfluidic chip 500 arranged in a row in sequence.

[0154] In addition, the optical detection module can further include a second heat dissipation channel 230 for dissipating heat of the optical detection assembly 210.

[0155] In one embodiment, as shown in FIGS. 16-21, the optical detection assembly 210 includes: an excitation light emitting unit capable of emitting excitation light to the microfluidic chip moved to the detection position; and a light signal receiving unit capable of receiving the fluorescence light signal from the microfluidic chip and converting the light signal into an electrical signal.

[0156] Through the above scheme, the excitation light emitting unit can emit excitation light to the microfluidic chip, so that the substance in the microfluidic chip emits fluorescence of a specific wavelength, and the light signal receiving unit receives the fluorescence of the specific wavelength and converts it into an electrical signal, thereby facilitating subsequent analysis and display (e.g., in the form of a two-dimensional digital signal). The process and required hardware devices for analysis and display are prior art, and will not be described herein.

[0157] Specifically, as to the excitation light emitting unit, it comprises: an excitation light emitting subunit 210-110 capable of emitting excitation light; a first light filtering subunit 210-120 provided with two or more (such as M, M is an integer greater than or equal to 2) first light filters 210-122, each of which is capable of filtering the excitation light emitted by the excitation light emitting subunit 210-110 to obtain excitation light of different wave bands respectively; and a first switching subunit 210-130 capable of moving and / or rotating the first light filtering subunit 210-120 to switch different first light filters 210-122 to the light path (the path between the excitation light emitting subunit 210-110 and the microfluidic chip) of the excitation light emitting subunit 210-110.

[0158] In the existing optical detection scheme, in order to meet the detection requirements, it is often necessary to set up multiple independent light emitting channels to obtain excitation light of different wave bands, but this scheme greatly affects the miniaturization and integration of the optical detection module. In the above scheme of the present application, only one excitation light emitting subunit 210-110 needs to be set up, which can emit light of a wide wave band in the excitation light range, and the first switching subunit 210-130 moves and / or rotates the first light filtering subunit 210-120 to switch different first light filters 210-122 to the light path of the excitation light emitting subunit 210-110, so that the excitation light emitting unit can emit excitation light of different wave bands to the microfluidic chip. Compared with the existing scheme, the above scheme of the present application has a simpler and more compact structure and is lower in cost.

[0159] As to the way in which the first switching subunit 210-130 moves and / or rotates the first light filtering subunit 210-120, the present application does not have specific limitations as long as it can respectively make different first light filters 210-122 located on the light path of the excitation light emitting subunit 210-110. Specifically, in the present embodiment, the switching of the first light filters 210-122 is realized by rotating the first light filtering subunit 210-120 driven by the first switching subunit 210-130. The first light filtering subunit 210-120 comprises a first light filtering wheel 210-121, a plurality of through holes are provided on the first light filtering wheel 210-121 along the circumferential direction of the first light filtering wheel 210-121, each first light filter 210-122 is arranged in each through hole, and the first switching subunit 210-130 is specifically a stepper motor which rotates the first light filtering wheel 210-121 by means of a synchronous toothed belt to realize the switching of each first light filter 210-122.

[0160] As to the excitation light emitting subunit 210-110, in the embodiment, it comprises an excitation light source 210-112 and a first light beam shaping lens group 210-112 arranged on the light path of the excitation light source 210-112. As described above, the excitation light source 210-112 can emit a wide range of wave bands in the excitation light range (preferably the visible light range), and the first light beam shaping lens group 210-112 can form a collimated uniform light beam from the light emitted by the excitation light source 210-112. As to the type of excitation light source, the present application does not have specific limitations, and those skilled in the art can make appropriate selection on the basis of existing excitation light sources as needed, such as selecting a white LED light source.

[0161] In order to ensure the normal operation of the excitation light source for a long time, a first heat dissipation member 210-140 can be arranged on the back of the excitation light source to ensure that the excitation light source is in a rated temperature state. The first heat dissipation member 210-140 can be a heat dissipation fin and / or a heat dissipation fan. In the embodiment, it is a heat dissipation fin arranged on the back of the excitation light source.

[0162] In the embodiment, the light signal receiving unit comprises a light signal receiving subunit (not shown in the figure), which can convert the fluorescent signal from the microfluidic chip into an electrical signal; a second light filtering subunit 210-220, which is provided with two or more second light filters 210-222, each of which can filter the excitation fluorescence from the microfluidic chip to obtain different wave bands of fluorescence; and a second switching subunit, which can drive the second light filtering subunit 210-220 to move and rotate, so as to switch different second light filters 210-222 to the receiving light path of the light signal receiving subunit.

[0163] As to the light signal receiving subunit, it can be a commonly used photosensitive chip (such as a CMOS sensor) in the art.

[0164] When a plurality of first light filters 210-122 and second light filters 210-222 are arranged respectively, those skilled in the art know that they need to be one-to-one corresponding during specific detection. That is, the first light filters 210-122 and the second light filters 210-222 are matched with the spectral characteristics of the used fluorescein, more specifically, the excitation light filtered by the first light filters 210-122 can excite the fluorescein to emit fluorescence of a specific wavelength, and the second light filters 210-222 only allow the fluorescence of the specific wavelength to pass, so as to be converted into an electrical signal by the light signal receiving subunit and can be analyzed, processed and displayed subsequently.

[0165] By the above scheme, only one optical signal receiving subunit needs to be provided, and by driving the second light filtering subunit 210-220 to move and / or rotate through the second switching subunit, different second filters can be switched on the receiving light path of the optical signal receiving subunit (the path between the optical signal receiving subunit and the microfluidic chip), so that the fluorescent signal emitted from the microfluidic chip can be filtered through the second filter, so that the optical signal receiving subunit can receive different wavebands of fluorescence, thereby achieving more abundant detection through a simple and compact structure.

[0166] As for the manner in which the second switching subunit drives the second light filtering subunit 210-220 to move and / or rotate, as long as it can enable different second filters 210-222 to be located on the receiving light path of the optical signal receiving subunit, the present application does not have specific limitations. Specifically, in the present embodiment, the second filters 210-222 are switched by driving the second light filtering subunit 210-220 to rotate through the second switching subunit. The second light filtering subunit 210-220 includes a second filter wheel 210-221, a plurality of through holes are provided on the second filter wheel 210-221 along the circumferential direction of the second filter wheel 210-221, each second filter 210-222 is provided in each through hole, and the second switching subunit (not shown in the figure) is specifically a stepper motor, which drives the second filter wheel 210-221 to rotate through a synchronous toothed belt or other existing manner, so as to switch each second filter 210-222.

[0167] In order to ensure the accuracy of detection, the optical signal receiving unit further comprises a light receiving subunit 210-230, which is arranged between the second light filtering subunit and the microfluidic chip, and the light receiving subunit 210-230 comprises a light shielding cylinder 210-231 and a second beam shaping lens group 210-232 arranged in the light shielding cylinder 210-231. Thus, the external stray light can be prevented from being received by the optical signal receiving subunit as much as possible, and the fluorescent light from the microfluidic chip can form a collimated uniform light beam and be imaged on the photosensitive surface of the subsequent photosensitive chip through the second beam shaping lens group 210-232.

[0168] [According to Rule 91 correction 15.07.2025] Further, as shown in Figures 19-21, the optical detection assembly further comprises a complementary color unit, which comprises a plurality of (e.g., N, N is an integer greater than or equal to 2) complementary color light emitting subunits 210-310, the complementary color light exciting unit can emit monochromatic light, and the wavebands of the monochromatic light emitted by different complementary color light emitting subunits 210-310 and the excitation light filtered by different first light filtering subunits are all different.

[0169] Specifically, the complementary light emitting subunit 210-310 includes a complementary light source 210-311, a third light beam shaping lens group 210-312 arranged on the light path (between the complementary light source 210-311 and the microfluidic chip) of the complementary light source 210-311, and a third filter 210-313.

[0170] Therefore, the complementary unit further supplements the wavelength band of the excitation light emitted by the excitation light emitting unit to the microfluidic chip, so as to emit more (M+N in total) wavelength bands of excitation light to the microfluidic chip to achieve better detection. The light emitted by the complementary light emitting subunit is not limited to visible light, and is preferably non-visible light bands, such as near-infrared light, far-infrared light, and / or near-ultraviolet light.

[0171] As for the type of complementary light source, the present application does not have specific restrictions, and those skilled in the art can make appropriate choices based on the existing excitation light source as needed, and preferably a single-color LED with good monochromaticity.

[0172] As for the parameters and number of lenses in the above light beam shaping lens groups (first light beam shaping lens group 210-112, second light beam shaping lens group 210-232, and third light beam shaping lens group 210-312), the present application does not have specific restrictions, as long as it can form a collimated uniform light beam.

[0173] Those skilled in the art know that the commonly used filters in the art include absorption filters and interference filters. Among them, the absorption filter utilizes the absorption difference of the material itself to different wavelengths of light, which can only allow certain wavelengths of light to pass through and absorb other wavelengths of light. The interference filter can utilize the interference principle of light, and the reflection interference between multiple optical films makes some wavelengths be enhanced to pass through, and other wavelengths of light are interfered and cancelled or reflected. As for the above filters (first filter, second filter, and second filter) of the present application, on the basis of the above scheme, those skilled in the art can select filters that allow the required wavelengths to pass through on the basis of existing filters.

[0174] As shown in FIGS. 16-21, the optical detection assembly 210 can further comprise a rack unit to set the specific structure of each unit of the optical detection assembly 210, so that the rack unit can be moved by the optical detection moving assembly 220 to move the optical detection assembly 210 as a whole. The rack unit can comprise a first fixed plate 210-410, which is arranged obliquely, and the excitation light emitting unit is fixed on the first fixed plate 210-410, so that the excitation light emitting unit can emit excitation light to the microfluidic chip from the side. The second fixed plate 210-420 is arranged horizontally, and the light signal receiving unit is fixed on the second fixed plate 210-420, so that the light signal receiving unit can receive the fluorescent signal from the microfluidic chip from the bottom of the microfluidic chip. As for the above-mentioned complementary unit, it can also be fixed obliquely on the detection rack unit to emit excitation light to the microfluidic chip from the side.

[0175] The optical detection assembly 210 can further comprise a housing unit (see FIG. 2), and other units of the optical detection assembly 210 are arranged in the housing unit, so as to prevent dust, moisture, pollution, and environmental light interference. Those skilled in the art know that necessary through holes should be provided on the housing unit to prevent blocking the light path.

[0176] In addition, in order to ensure the heat dissipation of the optical detection assembly 210 after the housing unit is arranged, the optical detection assembly 210 can further comprise a second heat dissipation member 210-500, which is a heat dissipation fan. The second heat dissipation member 210-500 can guide the heat of the optical detection assembly 210 out of the second heat dissipation channel 230. Since the optical detection assembly 210 is moved by the optical detection moving assembly 220, the second heat dissipation channel 230 is preferably telescopic along its axis (for example, the second heat dissipation channel 230 is a corrugated tube), and the two ends of the second channel are open, one end of which is fixed on the housing unit, and the other end extends out of the digital PCR all-in-one machine, so that the second heat dissipation member 210-500 guides the heat in the optical detection assembly 210 out through the second heat dissipation channel 230.

[0177] In one embodiment, as shown in FIGS. 1-3 and 5, the pressing plate module 300 further comprises:

[0178] The first pressing plate moving assembly can move the gas supply assembly 310 to and from the upper part of the temperature control module 100, and in this embodiment, it moves in the horizontal direction, and more specifically, it can move between the temperature control module 100 and the optical detection module 200.

[0179] Specifically, in the present embodiment, the first pressing plate moving assembly comprises:

[0180] a bearing frame 321 capable of bearing the gas supply assembly 310;

[0181] a first guide rail 322 arranged on the rack (in the present embodiment, two first guide rails 322 are arranged on the two sides inside the rack 900, respectively);

[0182] a sliding block 323 arranged at the bottom of the bearing frame 321;

[0183] a first movement unit 324, the movement end of which is directly or indirectly connected with the sliding block 323, so as to drive the gas supply assembly 310 arranged on the bearing frame 321 to slide along the first guide rail 322 through the sliding block 323.

[0184] As for the first movement unit, the present application does not have specific limitations. For example, a cylinder (as known to those skilled in the art, the push rod of the cylinder is its movement end, and the cylinder body is its fixed end), a lead screw (as known to those skilled in the art, the motor of the lead screw can be regarded as its fixed end, and the nut arranged on the screw rod can be regarded as its movement end) can be listed. As shown in FIG. 3, in the present embodiment, a lead screw is used, specifically, a motor (second motor 325) drives the screw rod (second screw rod 326) of the lead screw to rotate through a belt, so as to drive the nut arranged on the screw rod (second screw rod 326) to move, and the nut is fixed on the bearing frame 321, thereby realizing the driving of the gas supply assembly 310 arranged on the bearing frame 321 to slide along the first guide rail 322 through the sliding block 323 by the first movement unit 324.

[0185] In one embodiment, as shown in FIG. 5, the pressing plate module 300 further comprises:

[0186] a second pressing plate moving assembly, which can drive the gas supply assembly 310 to move to the gas supply position of the microfluidic chip 500 (i.e., the position capable of providing air pressure to the microfluidic chip 500) or move out of the gas supply position of the microfluidic chip 500 (in the present embodiment, the second pressing plate moving assembly specifically drives the gas supply assembly 310 to move in the vertical direction (up and down direction) to move to or move out of the gas supply position) when the first pressing plate moving assembly drives the gas supply assembly 310 to move to the upper part of the temperature control module 100.

[0187] The second pressing plate moving assembly comprises:

[0188] A second moving unit 331, a moving end of the second moving unit 331 is directly or indirectly connected with the air supply assembly 310 to drive the air supply assembly to move to or out of the air supply position of the microfluidic chip.

[0189] In order to facilitate the arrangement of the second pressing plate moving assembly, in the embodiment, the carrier frame 321 comprises a fixed plate 327, a fixed end of the second moving unit 331 is connected with the fixed plate, and a guide column 328, the guide column connects the sliding block with the fixed plate.

[0190] The second pressing plate moving assembly further comprises a movable plate 332, the movable plate 332 is provided with a guide hole, the guide hole is sleeved on the guide column 328, and the movable plate 332 can slide along the guide column 328; the air supply assembly 310 is fixed at the bottom of the movable plate 332; the fixed end of the second moving unit is connected with the fixed plate 327; the moving end of the second moving unit 331 is connected with the movable plate (that is, in the embodiment, the moving end of the second moving unit is indirectly connected with the air supply assembly 310 through the movable plate 332).

[0191] As for the second moving unit 331, the application does not have specific limitations, as long as it can drive the air supply assembly to move to or out of the air supply position of the microfluidic chip. Specifically, it can be listed, for example, a pneumatic cylinder (the push rod of the pneumatic cylinder is its moving end, and the cylinder body of the pneumatic cylinder is its fixed end), a lead screw (the motor of the lead screw can be regarded as its fixed end, and the nut provided on the screw rod of the lead screw can be regarded as its moving end). As shown in FIG. 3, in the embodiment, a lead screw is used, specifically, the (two) motors provided on the upper part of the fixed plate (third motor 333) drive the vertically arranged screw rod (third screw rod 334) to rotate, so that the movable plate 332 can move up and down, thereby driving the air supply assembly 310 fixed at the bottom of the movable plate 332 to move to or out of the air supply position of the microfluidic chip 500.

[0192] In one embodiment, as shown in FIGS. 9-11, the pressing plate module 300 further comprises:

[0193] A second heating assembly 340, when the second pressing plate moving assembly drives the air supply assembly 310 to move to the air supply position of the microfluidic chip 500, the second heating assembly 340 can heat the opposite side of the temperature control module 100 of the microfluidic chip 500 (in the embodiment, the upper surface of the microfluidic chip 500).

[0194] In the current PCR heating process (such as plate heating), the heating module is generally located only below the microfluidic chip, and thus the heat is conducted from below to the chip chamber or from above to the chip chamber, and the heat is quickly lost from above the chip, so that the chip cannot quickly reach or maintain the set temperature. In the embodiment, a second heating assembly is arranged above the chip and in contact with the chip during droplet generation and PCR reaction.

[0195] Specifically, the upper end of the second heating assembly 340 is connected with an elastic member 341 (a spring in the embodiment), and the elastic member 341 is directly or indirectly (indirectly through the movable plate 332 in the embodiment) connected with the moving end of the second moving unit 331 (that is, in the embodiment, the gas supply assembly 310 and the elastic member 341 for connecting the second heating assembly 340 are arranged on the movable plate 332); the gas outlets (the gas outlets connected with the chip inlets and outlets) of the gas supply assembly 310 are located on both sides of the second heating assembly 340, that is, the second heating assembly 340 is located between the gas outlets (the gas outlets connected with the chip inlets and outlets). Thus, the gas outlets of the gas supply assembly 310 located on both sides can output or stop outputting air pressure to the inlets and outlets on both sides of the microfluidic chip 500, and the second heating assembly 340 located in the middle can heat the reaction area located in the middle of the microfluidic chip 500.

[0196] The second heating assembly 340 includes one or more (such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20 or more) second heating units 342; each second heating unit 342 corresponds to a reaction area of the microfluidic chip 500. Thus, each microfluidic chip 500 can be independently temperature-controlled to perform PCR reaction under different temperature conditions.

[0197] As for the second temperature control unit 342, the present application does not have specific limitations as long as it can heat the microfluidic chip. In the embodiment, the second temperature control unit 342 is an electric heater.

[0198] Further, as shown in FIGS. 10 and 11, the second heating unit 342 is provided with a plurality of blind holes 343 facing the reaction area of the microfluidic chip, thereby constituting a heat preservation mechanism, greatly reducing the heat conduction such as heat convection, achieving heat preservation on the surface of the chip, making it closer to the temperature of the semiconductor cooler on the lower surface of the chip, ensuring that the required temperature is maintained in the chip, and achieving more accurate control of the droplet generation temperature and the PCR temperature.

[0199] In addition, the second temperature control unit 342 can also be a pressure plate with heat insulation material or heat insulation structure, which can quickly adjust the temperature or maintain the set temperature in the chip chamber by reducing heat dissipation above the microfluidic chip.

[0200] In one embodiment, as shown in Figures 12-14, the pressing plate module 300 is provided with oppositely arranged limiting grooves 350. Specifically, in the present embodiment, the limiting grooves 350 are respectively arranged on the two vertically arranged plates.

[0201] The pressing plate module 300 further comprises:

[0202] a baffle 360, which comprises:

[0203] a baffle body 361;

[0204] first protrusions 362 oppositely arranged on both sides of the baffle body 361, the baffle body 361 being located inside the oppositely arranged limiting grooves 350 and the first protrusions 362 being respectively located inside the limiting grooves 350;

[0205] second protrusions 363 arranged at the bottom of the baffle body 361;

[0206] When the chip moving module 400 moves the microfluidic chip 500 to the detection position of the optical detection module 200, the baffle 360 separates the microfluidic chip.

[0207] During optical detection, excitation light needs to enter the chip from the bottom of the chip or above the chip. If the excitation light enters the chip from the side of the chip, noise will be generated, affecting the scanning detection.

[0208] When using the digital PCR integrated machine of the present application, when the microfluidic chip is heated for PCR reaction, as shown in Figure 13, due to the support of the second protrusions 363 below the baffle body 361 on the surface of the temperature control module, only the second protrusions 363 below the baffle are in contact with the surface of the temperature control module 100, which ensures that the contact area between the baffle and the temperature control module 100 is very small, limited to the second protrusions 363, thereby greatly reducing the heat loss through the baffle. When optical detection is performed, as shown in Figure 14, the microfluidic chip 500 is moved to the detection position above the optical detection module 200. At this time, there is no object supporting the two second protrusions 363 below the baffle body 361. Due to the action of gravity, the baffle will fall a certain distance. At this time, the side of the chip is blocked by the baffle body 361, preventing the excitation light from entering from the side of the chip, thereby ensuring the accuracy of optical detection.

[0209] In one embodiment, as shown in FIG. 12, the pressing plate module 300 further comprises an elastic assembly 370, which is directly or indirectly fixed to the bottom of the movable plate 332, and when the air supply assembly 310 is at the air supply position of the microfluidic chip 500, the bottom of the elastic assembly 370 is at the top of the microfluidic chip 500 or the tray 600 for loading the microfluidic chip and is in a compressed state.

[0210] Since the microfluidic chip 500 is heated and pressed by the pressing plate module 300 (such as the air supply assembly 310 and the second heating unit 342) during PCR, it is easy to stick to the bottom of the pressing plate module 300 (such as the air supply assembly 310 and the second heating unit 342). After setting the elastic assembly 370 of the present embodiment, when the air supply assembly 310 is away from the air supply position of the microfluidic chip 500, the elastic assembly pushes the microfluidic chip 500 away from the bottom of the air supply assembly 310, thereby preventing the microfluidic chip 500 from sticking to the bottom of the pressing plate module, facilitating the automatic replacement of the microfluidic chip 500 by the digital PCR integrated machine of the present application.

[0211] Specifically, in the present embodiment, as shown in FIG. 12, the elastic assembly 370 comprises a plurality of elastic units, which comprise compression springs and movable columns arranged in vertical through holes, the top and bottom of the movable column are provided with protrusions, and the compression spring is arranged between the lower part of the through hole and the protrusion at the bottom of the movable column.

[0212] In one embodiment, the air supply assembly 310 can move between the loading position of the temperature control module 100 and the detection position of the optical detection module 200 (such as by the first pressing plate moving assembly and the second pressing plate moving assembly described above); the chip moving module 400 is directly or indirectly fixedly connected to the air supply assembly 310 (in the present embodiment, the air supply assembly 310 and the chip moving module 400 are both fixed to the movable plate 332, thereby achieving indirect fixation).

[0213] In the present embodiment, the chip moving module 400 is fixed to the air supply assembly 310, and when the chip moving module 400 fixes the microfluidic chip or the tray 600 for loading the microfluidic chip, the air supply assembly 310 can move between the loading position of the temperature control module 100 and the optical detection module 200, and the microfluidic chip 500 can be moved from the temperature control module 100 to the detection position of the optical detection module 200 by the chip moving module 400, thereby facilitating the detection of the microfluidic chip by the optical detection module. That is, in the present embodiment, the chip moving module 400 drives the microfluidic chip 500 to move from the loading position of the temperature control module 100 to the detection position of the optical detection module 200 by the air supply assembly 310 moving between the temperature control module 100 and the optical detection module 200.

[0214] Of course, a chip moving module completely independent of the pressing plate module can also be provided, which independently drives the microfluidic chip 500 from the temperature control module 100 to the detection position of the optical detection module 200 after the reagent in the microfluidic chip 500 completes PCR. Compared with the scheme of the present embodiment, the miniaturization and simplification of the device are facilitated.

[0215] The way in which the chip moving module 400 fixes the microfluidic chip 500 or the tray 600 for loading the microfluidic chip is not specifically limited, and can be achieved by structures such as magnetic adsorption or jaw fixation. In the present embodiment, the chip moving module 400 comprises a chip holder assembly, one end of the chip holder assembly is directly or indirectly fixedly connected with the air supply assembly 310 (in the present embodiment, the air supply assembly 310 is indirectly fixedly connected with the movable plate 332), and the other end extends to the lower part of the air supply assembly 310, so that when the second pressing plate moving assembly drives the air supply assembly to move out of the air supply position of the microfluidic chip, the chip holder assembly can lift the tray 600 for loading the microfluidic chip, so that the microfluidic chip leaves the bearing position of the temperature control module.

[0216] In one embodiment of the chip moving module 400 of the present embodiment, as shown in FIGS. 5 and 6, the chip holder assembly comprises: a chip holder body 410, one end of the chip holder body 410 is directly or indirectly fixedly connected with the air supply assembly 310 (in the present embodiment, the air supply assembly 310 is indirectly fixedly connected with the movable plate 332); a third movement unit 420 arranged on the chip holder body; and a jaw 430 arranged at the movement end of the third movement unit 420, which can drive the jaw to clamp the tray 600 for loading the microfluidic chip when the air supply assembly is located at the air supply position of the microfluidic chip. In the present embodiment, the tray 600 is provided with grooves on both sides, and when the movement end of the third movement unit 420 drives the jaw 430 to clamp the microfluidic chip, the jaw 430 is deep into the grooves, so that when the second movement unit 331 drives the movable plate 332 to move upward, the chip moving module 400 can drive the microfluidic chip 500 to move out of the bearing position of the temperature control module 100, and then the air supply assembly 310 and the microfluidic chip 500 can be driven together to move to the optical detection module by the first pressing plate moving assembly.

[0217] As to the third moving unit 420, the present application is not specifically limited as long as it can drive the clamping jaw 430 to clamp or release the microfluidic chip 500 or the tray 600 for loading the microfluidic chip. Specifically, it can be listed that, for example, a pneumatic cylinder (the push rod of the pneumatic cylinder is the moving end thereof, and the cylinder body of the pneumatic cylinder is the fixed end thereof), a lead screw (the motor of the lead screw can be regarded as the fixed end thereof, and the nut arranged on the screw rod of the lead screw can be regarded as the moving end thereof). As shown in FIG. 6, in the present embodiment, the oppositely arranged two lead screws respectively drive the oppositely arranged clamping jaws 430 to move towards each other or move away from each other, so as to clamp or release the microfluidic chip 500 or the tray 600 for loading the microfluidic chip. Thus, when the second pressure plate moving assembly drives the gas supply assembly to move out of the gas supply position and the first pressure plate moving assembly drives the gas supply assembly to move to the optical detection module, the tray 600 can be lifted by the chip carrier unit 440, and meanwhile, the tray 600 can be tightly fixed by the chip moving module (the clamping jaw 430 clamps the tray 600) to prevent the tray 600 from moving relative to the chip moving module in the vertical direction and the horizontal direction, so as to increase the reliability of the equipment operation and the accuracy of the detection result.

[0218] In another embodiment of the chip moving module 400 of the present embodiment, as shown in FIG. 7 and FIG. 8, the chip carrier assembly comprises: a pair of oppositely arranged chip carrier units 440, one end of the chip carrier unit 440 is directly or indirectly fixedly connected with the gas supply assembly 310 (in the present embodiment, it is fixedly connected on the movable plate 332, and indirectly connected with the gas supply assembly 310), and the other end of the chip carrier unit 440 extends to the lower part of the gas supply assembly 310; and a third protrusion 450 arranged on the upper surface of the other end of the chip carrier unit 440. At this time, the third protrusion 450 corresponds to the through hole arranged on the tray 600 or the blind hole arranged on the bottom surface, so that when the second pressure plate moving assembly drives the gas supply assembly to move out of the gas supply position and the first pressure plate moving assembly drives the gas supply assembly to move to the optical detection module, the tray 600 can be lifted by the chip carrier unit 440, and meanwhile, the tray 600 can be prevented from moving relative to the chip moving module in the horizontal direction by the third protrusion 450 corresponding to the through hole or the blind hole arranged on the tray 600, so as to increase the reliability of the equipment operation and the accuracy of the detection result.

[0219] As known by those skilled in the art, the vertical distance between the upper part of the third protrusion 450 and the lower part of the gas supply assembly 310 should be greater than the thickness of the tray 600 used, so that the tray 600 can be accommodated therebetween. In the present application, the "vertical distance" refers to the distance between the vertical feet of the vertical lines passing through the two points, respectively.

[0220] In addition, the digital PCR all-in-one machine of the present application can further be provided with a display 800, which can be a touch display, for example, to serve as the input of instructions and the output of detection results, etc.

[0221] The digital PCR all-in-one machine of the present application can further comprise a control module 700 to control the above modules. Specifically, the control module 700 is electrically connected to each motor (which can be a stepper motor), heating element, photosensitive chip, and / or display in each of the above modules for automatic control. As for the control module, a module using a single-chip microcomputer for control or a PLC control module can be used. Based on the above technical solutions given in the present application, those skilled in the art are aware of how to achieve the electrical connection of the control module 700 to each motor, heating unit, photosensitive chip, and / or display and control according to the prior art, which will not be described here in detail.

[0222] Although the embodiments of the present application are described above, the present application is not limited to the specific embodiments and application fields described above, and the specific embodiments described above are only illustrative and instructive, but not limiting. Those skilled in the art can make many forms under the guidance of the present application and without departing from the scope protected by the claims of the present application, which all belong to the present application claimed.

Claims

1. A digital PCR all-in-one machine, wherein, The device comprises: a temperature control module, a carrying position of which is capable of carrying a microfluidic chip or a tray for loading the microfluidic chip, and the temperature control module is capable of controlling the temperature of a reaction area of the microfluidic chip located in the carrying position of the temperature control module to perform a PCR reaction; an optical detection module, which is arranged on one side of the temperature control module and is capable of detecting the PCR reaction result in the reaction area of the microfluidic chip; a pressing plate module, which comprises a gas supply assembly, the gas supply assembly is capable of moving to and out of the upper part of the temperature control module, and when the gas supply assembly moves to the upper part of the temperature control module, the gas supply assembly is capable of outputting or stopping outputting air pressure to the inlet of the microfluidic chip located in the carrying position of the temperature control module; a chip moving module, which is capable of moving the microfluidic chip from the carrying position of the temperature control module to a detection position of the optical detection module, so that the optical detection module can detect the microfluidic chip.

2. The digital PCR all-in-one machine according to claim 1, wherein the temperature control module comprises a first temperature control assembly, which is capable of controlling the temperature of at least one surface of the microfluidic chip located in the carrying position of the temperature control module.

3. The digital PCR all-in-one machine according to claim 2, wherein the first temperature control assembly comprises one or more first temperature control units, each of which corresponds to a reaction area of the microfluidic chip.

4. The digital PCR all-in-one machine according to claim 2, wherein the temperature control module further comprises a first heat dissipation assembly; the first heat dissipation assembly comprises: a first heat dissipation channel located at the bottom of the first temperature control assembly; and a heat dissipation fan arranged in the first heat dissipation channel.

5. The digital PCR all-in-one machine according to claim 1, wherein the optical detection module comprises: an optical detection assembly, which is capable of detecting the PCR reaction result in the reaction area of the microfluidic chip; an optical detection moving assembly, which is capable of moving the optical detection assembly to detect the PCR reaction result in different microfluidic chips and / or different reaction areas of a microfluidic chip.

6. The digital PCR all-in-one machine according to claim 5, wherein the optical detection assembly comprises: an excitation light emitting unit, which is capable of emitting excitation light to the microfluidic chip moved to the detection position; a light signal receiving unit, which is capable of receiving fluorescence from the microfluidic chip and converting the fluorescence into an electrical signal.

7. The digital PCR all-in-one machine according to claim 6, wherein the excitation light emitting unit comprises: an excitation light emitting subunit, which is capable of emitting excitation light; a first light filtering subunit, on which two or more first light filters are arranged, each of which is capable of filtering the excitation light emitted by the excitation light emitting subunit to obtain excitation light of different wave bands, respectively. The first switching subunit can drive the first filter subunit to move and / or rotate, so as to switch different first filters to the light path of the excitation light emitting subunit.

8. The digital PCR all-in-one machine of claim 7, wherein, The excitation light emitting subunit comprises an excitation light source and a first light beam shaping lens group arranged on the light path of the excitation light source.

9. The digital PCR all-in-one machine of claim 6, wherein, The light signal receiving unit comprises: The light signal receiving subunit can convert the fluorescence from the microfluidic chip into an electrical signal; The second filter subunit is provided with two or more second filters, and each second filter can filter the fluorescence from the microfluidic chip to obtain fluorescence of different wave bands, respectively; The second switching subunit can drive the second filter subunit to move and / or rotate, so as to switch different second filters to the receiving light path of the light signal receiving subunit.

10. The digital PCR all-in-one machine of claim 9, wherein, The light signal receiving unit further comprises: The light receiving subunit is arranged between the second filter subunit and the microfluidic chip, and comprises a light shielding cylinder and a second light beam shaping lens group arranged in the light shielding cylinder.

11. The digital PCR all-in-one machine of claim 7, wherein, The optical detection assembly further comprises: The complementary color unit comprises a plurality of complementary color light emitting subunits, the complementary color light emitting subunits can emit monochromatic light, and the wave bands of the monochromatic light emitted by different complementary color light emitting subunits and the excitation light filtered by different first filter subunits are all different.

12. The digital PCR all-in-one machine of claim 11, wherein, The complementary color light emitting subunit comprises a complementary color light source, a third light beam shaping lens group arranged on the light path of the complementary color light source, and a third filter.

13. The digital PCR all-in-one machine of claim 1, wherein, When the gas supply assembly moves to the upper part of the temperature control module, the gas supply assembly can also output or stop outputting air pressure to the outlet of the microfluidic chip located in the temperature control module carrying position.

14. The digital PCR all-in-one machine of claim 1, wherein, The pressure plate module further comprises: The first pressure plate moving assembly can drive the gas supply assembly to move to the upper part of the temperature control module and move out of the upper part of the temperature control module.

15. The digital PCR all-in-one machine of claim 14, wherein, The first pressure plate moving assembly comprises: The carrying frame can carry the gas supply assembly; The first guide rail is arranged on the rack; The slider is arranged at the bottom of the carrying frame; The movement end of the first movement unit is directly or indirectly connected with the slider, so as to drive the gas supply assembly arranged on the carrying frame to slide along the first guide rail through the slider. 16.The digital PCR machine of claim 15, wherein, The pressing plate module further comprises: A second pressing plate moving assembly, when the first pressing plate moving assembly drives the gas supply assembly to move to the upper portion of the temperature control module, the second pressing plate moving assembly can drive the gas supply assembly to move to the gas supply position of the microfluidic chip or move out of the gas supply position of the microfluidic chip. 17.The digital PCR machine of claim 16, wherein, The second pressing plate moving assembly comprises: A second moving unit, a moving end of the second moving unit is directly or indirectly connected with the gas supply assembly to drive the gas supply assembly to move to the gas supply position of the microfluidic chip or move out of the gas supply position of the microfluidic chip. 18.The digital PCR machine of claim 17, wherein, The bearing frame comprises: A fixed plate; and A guide column connecting the sliding block with the fixed plate; The second pressing plate moving assembly further comprises: A movable plate, the movable plate is provided with a guide hole, the guide hole is sleeved on the guide column, and the movable plate can slide along the guide column; a fixed end of the second moving unit is connected with the fixed plate; a moving end of the second moving unit is connected with the movable plate; and the gas supply assembly is fixed to the bottom of the movable plate. 19.The digital PCR machine of claim 17, wherein, The pressing plate module further comprises: A second heating assembly, when the second pressing plate moving assembly drives the gas supply assembly to move to the gas supply position of the microfluidic chip, the second heating assembly can heat the opposite side of the temperature control module of the microfluidic chip. 20.The digital PCR machine of claim 19, wherein, An upper end of the second heating assembly is connected with an elastic member, and the elastic member is directly or indirectly fixedly connected with the moving end of the second moving unit; Gas outlets of the gas supply assembly are located on both sides of the second heating assembly. 21.The digital PCR machine of claim 19, wherein, The second heating assembly comprises more than one second heating unit; Each second heating unit corresponds to a reaction area of the microfluidic chip. 22.The digital PCR machine of claim 21, wherein, The second heating unit is provided with a plurality of blind holes facing the reaction area of the microfluidic chip. 23.The digital PCR machine of claim 1, wherein, The pressing plate module is provided with two oppositely arranged limiting grooves; The pressing plate module further comprises a baffle, and the baffle comprises: A baffle body, the baffle body is located on the inner side of the oppositely arranged limiting grooves; More than two first protrusions, the more than two first protrusions are arranged on both sides of the baffle body, and the first protrusions are respectively located in the limiting grooves; More than two second protrusions, the more than two second protrusions are arranged on the bottom of the baffle body; When the chip moving module moves the microfluidic chip to the detection position of the optical detection module, the baffle separates the microfluidic chip. 24.The digital PCR machine of claim 18, wherein, The pressing plate module further comprises: An elastic assembly is directly or indirectly fixed to the bottom of the movable plate, and the bottom of the elastic assembly is located at the top of the microfluidic chip or a tray for loading the microfluidic chip and is in a pressed state when the air supply assembly is located at the air supply position of the microfluidic chip.

25. The digital PCR all-in-one machine of claim 16, wherein, The air supply assembly is capable of moving between the bearing position of the temperature control module and the detection position of the optical detection module. The chip moving module is directly or indirectly fixedly connected with the air supply assembly.

26. The digital PCR all-in-one machine of claim 25, wherein, The chip moving module comprises: A chip carrier assembly, one end of which is directly or indirectly fixedly connected with the air supply assembly, and the other end extends to the lower part of the air supply assembly, so that the chip carrier assembly can lift a tray for loading the microfluidic chip when the air supply assembly is moved out of the air supply position of the microfluidic chip by the second pressing plate moving assembly, and the microfluidic chip leaves the bearing position of the temperature control module.

27. The digital PCR all-in-one machine of claim 26, wherein, The chip carrier assembly at least comprises: A pair of chip carrier bodies, one end of which is directly or indirectly fixedly connected with the air supply assembly; A third movement unit, which is arranged on the chip carrier body; A clamping jaw, which is arranged at the movement end of the third movement unit, and the movement end of the third movement unit can drive the clamping jaw to clamp the tray for loading the microfluidic chip when the air supply assembly is located at the air supply position of the microfluidic chip.

28. The digital PCR all-in-one machine of claim 26, wherein, The chip carrier assembly comprises: A pair of chip carrier units, one end of which is directly or indirectly fixedly connected with the air supply assembly, and the other end of the chip carrier unit extends to the lower part of the air supply assembly.

29. The digital PCR all-in-one machine of claim 28, wherein, The chip carrier assembly further comprises: A third protrusion, which is arranged on the upper surface of the other end of the chip carrier unit.

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