Digital PCR all-in-one machine

Abstract

The utility model provides a digital PCR (polymerase chain reaction) all-in-one machine, which comprises a temperature control module capable of controlling the temperature of a reaction area of a micro-fluidic chip positioned at a bearing position of the temperature control module so as to carry out PCR reaction; the optical detection module can be used for detecting a PCR reaction result in the reaction area of the micro-fluidic chip; the pressing plate module comprises an air supply assembly, and when the air supply assembly moves to the upper portion of the temperature control module, the air supply assembly can output air pressure to an inlet of the micro-fluidic chip located at the bearing position of the temperature control module or stop outputting the air pressure; and the chip moving module can move the micro-fluidic chip from the bearing of the temperature control module to the detection position of the optical detection module. According to the digital PCR all-in-one machine provided by the invention, an instrument can automatically complete droplet generation, PCR reaction and optical scanning detection only by adding a sample into the micro-fluidic chip and loading the chip into the digital PCR all-in-one machine, so that the convenience degree of the instrument in use is greatly increased.

Description

Technical Field

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

[0002] Microfluidics is a technology for precisely controlling and manipulating microscale fluids. It has the characteristics of small capacity (nanoliter, picoliter, femtoliter level) and low energy consumption. People use microfluidics to integrate basic operating units such as sample preparation, reaction, separation, and detection in biological, chemical, and medical analysis processes into microfluidic chips with microfluidic pipelines as the basic structure, achieving advantages such as rapid sample processing and detection, and low reagent and sample usage. Due to its huge potential in the fields of biology, chemistry, and medicine, it has developed into a new research field that intersects biology, chemistry, medicine, fluids, electronics, materials, and mechanics.

[0003] Digital PCR, a method of nucleic acid quantification based on single-molecule PCR counting, is an absolute quantitative method. The principle is to disperse a large amount of diluted nucleic acid solution into the microreactor or droplet of the microfluidic chip through microfluidics or dropletization, and the number of nucleic acid templates in each microreactor is less than or equal to 1. After the PCR cycle, the reactor or microdroplet with a nucleic acid molecule template will give a fluorescent signal, and the reactor or microdroplet without a template will not have a fluorescent signal. According to the relative proportion and the volume of the reactor, the nucleic acid concentration of the original solution is calculated.

[0004] In the existing digital PCR system, the user uses a pipette to transfer the generated droplets to a PCR reaction tube, and then places the tube into a PCR thermal cycler for PCR reaction. After the reaction is completed, the user takes out the PCR tube and places it into a droplet fluorescence detector for droplet fluorescence detection. The detection principle is to inject droplets and oil into a capillary tube, and the droplets pass through the detection position individually, and the instrument reads whether the droplets have fluorescence. However, the system has the following disadvantages: (1) After the system generates droplets, the user needs to manually transfer the droplets to a 96-well plate before the next PCR reaction can be carried out, which will cause the loss or breakage of the droplets; (2) Due to the limitation of heat transfer rate (the sample volume in the test tube is thick, and heating starts from the outside of the test tube), the PCR reaction speed in the test tube is slow, so the entire PCR process takes a long time, generally requiring 150 minutes; (3) After the droplets pass through the detection position individually, they enter the waste liquid bottle, and the sample cannot be traced back.

[0005] There is currently no automated PCR instrument on the market that is specifically designed for microfluidic chips. Utility Model Content

[0006] In order to solve the problems in the prior art, the present application provides a digital PCR all-in-one machine. The technical solution of the present application is as follows:

[0007] The present application provides a digital PCR all-in-one machine, comprising:

[0008] A temperature control module, wherein the bearing position of the temperature control module can bear a microfluidic chip or a tray for bearing the microfluidic chip, and the temperature control module can control the temperature of a reaction area of ​​the microfluidic chip located at the bearing position of the temperature control module to perform a PCR reaction;

[0009] An optical detection module, which is disposed on one side of the temperature control module and is capable of detecting a PCR reaction result in a reaction area of ​​the microfluidic chip;

[0010] A pressing plate module, the pressing plate module comprising an air supply component, the air supply component can be moved to the upper part of the temperature control module and moved out from the upper part of the temperature control module; when the air supply component moves to the upper part of the temperature control module, the air supply component can output air pressure to the inlet of the microfluidic chip located at the bearing position of the temperature control module or stop outputting air pressure;

[0011] A chip moving module, wherein the chip moving module can move the microfluidic chip from the bearing position of the temperature control module to the detection position of the optical detection module, so that the optical detection module can detect the microfluidic chip.

[0012] Furthermore, the temperature control module includes a first temperature control component, and the first temperature control component can control the temperature of at least one surface of the microfluidic chip located at the bearing position of the temperature control module.

[0013] Furthermore, the first temperature control component includes more than one first temperature control unit, and each of the first temperature control units corresponds to a reaction area of ​​the microfluidic chip.

[0014] Furthermore, the temperature control module also includes a first heat dissipation component; the first heat dissipation component includes: a first heat dissipation channel, the first heat dissipation channel is located at the bottom of the first temperature control component; and a heat dissipation fan, the heat dissipation fan is arranged in the first heat dissipation channel.

[0015] Furthermore, the optical detection module includes: an optical detection component, which can detect the PCR reaction results in the reaction area of ​​the microfluidic chip; and an optical detection moving component, which can drive the optical detection component to move to detect the PCR reaction results in different microfluidic chips and / or different reaction areas of the microfluidic chip.

[0016] Furthermore, when the air supply component moves to the upper part of the temperature control module, the air supply component can also output air pressure to the outlet of the microfluidic chip located at the bearing position of the temperature control module or stop outputting air pressure.

[0017] Furthermore, the pressure plate module also includes: a first pressure plate moving assembly, which can drive the air supply assembly to move to the upper part of the temperature control module and move out from the upper part of the temperature control module.

[0018] Furthermore, the first pressure plate moving assembly includes: a carrier frame, which is capable of carrying the air supply assembly; a first guide rail, which is arranged on the frame; a slider, which is arranged at the bottom of the carrier frame; and a first motion unit, the motion end of the first motion unit is directly or indirectly connected to the slider to drive the air supply assembly arranged on the carrier frame to slide along the first guide rail through the slider.

[0019] Furthermore, the pressure plate module also includes: a second pressure plate moving component. When the first pressure plate moving component drives the gas supply component to move to the upper part of the temperature control module, the second pressure plate moving component can drive the gas supply component to move to the gas supply position of the microfluidic chip or move out of the gas supply position of the microfluidic chip.

[0020] Furthermore, the second pressure plate moving assembly includes: a second motion unit, a motion end of which is directly or indirectly connected to 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.

[0021] Furthermore, the carrier frame includes: a fixed plate; and a guide column, which connects the slider and the fixed plate; the second pressure plate moving assembly also includes: a movable plate, which is provided with a guide hole, which is sleeved on the guide column, and the movable plate can slide along the guide column; the fixed end of the second motion unit is connected to the fixed plate; the moving end of the second motion unit is connected to the movable plate; and the air supply assembly is fixed to the bottom of the movable plate.

[0022] Furthermore, the pressure plate module also includes: a second heating component. When the second pressure plate moving component drives the gas supply component to move to the gas supply position of the microfluidic chip, the second heating component can heat the opposite side of the temperature control module of the microfluidic chip.

[0023] Furthermore, an elastic member is connected to the upper end of the second heating component, and the elastic member is directly or indirectly fixedly connected to the moving end of the second moving unit; and the air outlet of the air supply component is located on both sides of the second heating component.

[0024] Furthermore, the second heating component includes more than one second heating unit; each of the second heating units corresponds to a reaction area of ​​the microfluidic chip.

[0025] Furthermore, the second heating unit is provided with a plurality of blind holes facing the reaction area of ​​the microfluidic chip.

[0026] Furthermore, the pressure plate module is provided with two relatively arranged limit grooves; the pressure plate module also includes a baffle, and the baffle includes: a baffle body, the baffle body is located on the inner side of the relatively arranged limit grooves; more than two first protrusions, more than two of the first protrusions are arranged on both sides of the baffle body, and the first protrusions are respectively located in the limit grooves; more than two second protrusions, more than two of the second protrusions are arranged at the bottom of the baffle body; 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.

[0027] Furthermore, the pressure plate module also includes: an elastic component, which is directly or indirectly fixed to the bottom of the movable plate, and when the air supply component is located at the air supply position of the microfluidic chip, the bottom of the elastic component is located at the top of the microfluidic chip or the tray for loading the microfluidic chip and is in a pressurized state.

[0028] Furthermore, the air supply component can move between the carrying position of the temperature control module and the detection position of the optical detection module; and the chip moving module is directly or indirectly fixedly connected to the air supply component.

[0029] Furthermore, the chip moving module includes: a chip holder assembly, one end of the chip holder assembly is directly or indirectly fixedly connected to the air supply assembly, and the other end extends to the lower part of the air supply assembly, so that when the second pressure 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 for loading the microfluidic chip, so that the microfluidic chip leaves the carrying position of the temperature control module.

[0030] Furthermore, the chip holder assembly at least includes: two chip holder bodies arranged in pairs, one end of the chip holder body being directly or indirectly fixedly connected to the gas supply assembly; the third motion unit, the third motion unit being arranged on the chip holder body; and a clamp, the clamp being arranged at a moving end of the third motion unit, and when the gas supply assembly is located at the gas supply position of the microfluidic chip, the moving end of the third motion unit can drive the clamp to clamp the tray for loading the microfluidic chip.

[0031] Furthermore, the chip bracket assembly includes: two chip bracket units arranged in pairs, one end of the chip bracket unit is directly or indirectly fixedly connected to the air supply assembly, and the other end of the chip bracket unit extends to the lower part of the air supply assembly.

[0032] Furthermore, the chip bracket assembly further includes: a third protrusion, and the third protrusion is arranged on the upper surface of the other end of the chip bracket unit.

[0033] Through the above-mentioned digital PCR all-in-one machine provided in the present application, it is only necessary to add the sample into the microfluidic chip and load the chip into the digital PCR all-in-one machine. The instrument can automatically complete droplet generation, PCR reaction and optical scanning detection, which greatly increases the convenience of using the instrument.

[0034] The above description is only an overview of the technical solution of the present application. In order to make the technical means of the present application clearer and to enable those skilled in the art to implement it according to the contents of the specification, and to make the above and other purposes, features and advantages of the present application more obvious and easy to understand, the specific implementation methods of the present application are exemplified below. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 : A schematic diagram of the side view structure of a digital PCR integrated machine from a first viewing angle in one embodiment of the present application;

[0036] Figure 2 : A schematic diagram of the side view structure of the digital PCR integrated machine from a second viewing angle in one embodiment of the present application;

[0037] Figure 3 : A schematic diagram of the side view structure of the digital PCR all-in-one machine from a third viewing angle in one embodiment of the present application;

[0038] Figure 4 : A schematic structural diagram of a cross section of a digital PCR integrated machine in one embodiment of the present application;

[0039] Figure 5 : A schematic diagram of the structure of the chip moving module in the digital PCR integrated machine in one embodiment of the present application;

[0040] Figure 6 : A schematic diagram of the structure of a chip moving module in a digital PCR all-in-one machine in one embodiment of the present application;

[0041] Figure 7 : A schematic diagram of the structure of the chip moving module in the digital PCR all-in-one machine in another embodiment of the present application;

[0042] Figure 8: A schematic diagram of the structure of a chip moving module in a digital PCR all-in-one machine in another embodiment of the present application;

[0043] Fig. 9 : A schematic diagram of a structure in which a gas supply component in a digital PCR integrated machine is located at a gas supply position of a microfluidic chip in one embodiment of the present application;

[0044] Fig.10 : A schematic diagram of the structure of the second temperature control unit in the digital PCR all-in-one machine in one embodiment of the present application;

[0045] Fig.11 : A bottom view schematic diagram of a second temperature control unit in a digital PCR all-in-one machine in one embodiment of the present application;

[0046] Fig.12 : A schematic diagram of the structure of the gas supply position where the gas supply component in the digital PCR integrated machine leaves the microfluidic chip in one embodiment of the present application;

[0047] Fig.13 : A schematic diagram of the position of the baffle when the gas supply assembly is located at the gas supply position of the microfluidic chip in one embodiment of the present application;

[0048] Fig.14 : A schematic diagram of the position of the baffle when the air supply assembly is located at the detection position of the optical detection module in one embodiment of the present application;

[0049] Fig.15 : Schematic diagram of the position of the microfluidic chip and the first temperature control unit in a digital PCR all-in-one machine in one embodiment of the present application.

[0050] Description of reference numerals:

[0051] 100, temperature control module; 110, first temperature control component; 111, first temperature control unit; 120, first heat dissipation component;

[0052] 200, optical detection module; 210, optical detection assembly; 220, optical detection moving assembly; 221, first motor; 222, first screw; 230, second heat dissipation channel;

[0053] 300, platen module; 310, air supply assembly; 321, carrier; 322, first guide rail; 323, slider; 324, first motion unit; 325, second motor; 326, second screw; 327, fixed plate; 328, guide column; 331, second motion unit; 332, movable plate; 333, third motor; 334, third screw; 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;

[0054] 400, chip moving module; 410, chip bracket body; 420, third moving unit; 430, clamping claw; 440, chip bracket unit; 450, third protrusion;

[0055] 500. Microfluidic chip;

[0056] 600, pallet;

[0057] 700, control module;

[0058] 800, display;

[0059] 900, rack. DETAILED DESCRIPTION

[0060] 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 other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present application are deemed to be equivalent replacement methods and fall within the scope of protection of the present application.

[0061] Those skilled in the art should understand that, in the disclosure of the present application, the terms "first", "second", "third", "fourth", "fifth", etc. are only used to distinguish different structures, but do not limit the number, connection relationship, etc. of specific structures; in addition, the directions or positional relationships indicated by "longitudinal", "transverse", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc. are based on the directions or positional relationships shown in the accompanying drawings, which are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation. Therefore, the above terms cannot be understood as limitations on the present application.

[0062] This embodiment provides a digital PCR all-in-one machine, such as Figure 1 to Figure 15 As shown, including:

[0063] A temperature control module 100, wherein the carrying position of the temperature control module 100 (i.e., the position for carrying the microfluidic chip 500 or the tray 600) can carry the microfluidic chip 500 or the tray 600 for carrying the microfluidic chip (specifically, in this embodiment, the carrying tray 600, on which the microfluidic chip 500 is loaded), and the temperature control module 100 can control the temperature of the reaction area (the area where the reaction chamber is set) of the microfluidic chip 500 located at the carrying position of the temperature control module 100 to perform a PCR reaction;

[0064] An optical detection module 200, which can detect the PCR reaction result in the reaction area of ​​the microfluidic chip 500;

[0065] A pressing plate module 300, wherein the pressing plate module 300 includes an air supply component 310, and the air supply component 310 can be moved to the upper part of the temperature control module 100 and moved out from the upper part of the temperature control module 100 (in this embodiment, specifically, moved in the horizontal direction); when the air supply component 310 moves to the upper part of the temperature control module 100, the air supply component 310 can output air pressure to the inlet of the microfluidic chip 500 located at the bearing position of the temperature control module 100 or stop outputting air pressure, so that the oil phase and the water phase in the microfluidic chip 500 can move from the inlet to the outlet of the microfluidic chip 500, so as to form droplets for PCR in the reaction chamber of the microfluidic chip 500;

[0066] The chip moving module 400 can move the microfluidic chip 500 from the bearing position of the temperature control module 100 to the detection position of the optical detection module 200 (i.e., the position where the optical detection module 200 can detect the microfluidic chip 500) (in this embodiment, specifically moves in the vertical direction), so as to facilitate the optical detection module 200 to detect the microfluidic chip 500.

[0067] Furthermore, in this embodiment, when the pressure plate module 300 moves to the upper part of the temperature control module 100, the gas supply component 310 can also output air pressure to the outlet of the microfluidic chip 500 located at the bearing position of the temperature control module 100 or stop outputting air pressure. Therefore, 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.

[0068] Specifically, in this embodiment, the gas supply component 310 is a gas supply pressure plate, and the gas supply pressure plate is provided with a gas flow channel inside, and the lower surface thereof is provided with a gas outlet corresponding to the inlet and outlet of the microfluidic chip, and it is provided with an air inlet hole connected to the gas source to provide gas of different pressures, so that the pneumatic pressure plate can provide different pressures to 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 characteristics of the flow channel wall during the droplet generation process, and is conducive to the generation of droplets of uniform and stable size; in the second step, a specific pressure is set for the oil phase and the water phase according to the size of the required droplets, and a specific size of droplets is generated or the size of the droplets is adjusted. The air supply pressure plate can provide different pressures to the inlet (oil phase and water phase) and outlet of the chip to control the movement of droplets, adjust the relative position of droplets in the chip chamber and the droplet density; it can also provide the same pressure to increase the boiling point of water and increase 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.

[0069] When using the digital PCR integrated machine provided in this embodiment, first, the pressing plate module 300 is not located on the upper part of the temperature control module 100, but is located on the upper part of the optical detection module 200 (such as Figure 1 , Figure 3 As shown in the position, the temperature control module 100 is exposed, and the microfluidic chip 500 loaded with the liquid phase and the oil phase is carried to the carrying position of the temperature control module 100 through the tray 600; then, the pressure plate module 300 is moved to the upper part of the temperature control module 100, and the gas supply component 310 outputs gas pressure to the inlet of the microfluidic chip 500 located at 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 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 a 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 uses the optical detection module 200 to detect the microfluidic chip 500.

[0070] By using the digital PCR all-in-one machine of the present application, one only needs to add the sample into the microfluidic chip and load the chip into the digital PCR all-in-one machine. The instrument can automatically complete droplet generation, PCR reaction and optical scanning detection, greatly increasing the convenience of using the instrument.

[0071] In one embodiment, Figure 1 , Fig.15As shown, the temperature control module 100 includes a first temperature control component 110 to perform temperature control on at least one surface of the microfluidic chip 500 located at the carrying position of the temperature control module 100 (the lower surface of the microfluidic chip 500 in this embodiment).

[0072] Specifically, in this embodiment, the first temperature control component 110 includes more than one (such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20 or more, 8 in this embodiment) first temperature control units 111, each of which corresponds to a reaction zone of the microfluidic chip 500, so that each microfluidic chip 500 can be independently tested. The independent first temperature control unit 111 can allow different temperatures to be set, such as forming a temperature gradient on the chip tested in parallel, to achieve a screening of the appropriate temperature required for the PCR reaction, or the screening and research of other temperature-sensitive conditions.

[0073] Regarding the first temperature control unit 111, the present application has no specific restrictions, 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, so that the temperature control can be performed more quickly and accurately, so as to facilitate the PCR reaction to be performed accurately under specific temperature conditions. In this embodiment, the first temperature control unit 111 is a TEC heating unit (semiconductor cooler, also called thermoelectric cooler), so as to be able to heat and cool the microfluidic chip 500.

[0074] Furthermore, in this embodiment, the temperature control module 100 further includes: a first heat dissipation component 120, the first heat dissipation component 120 including a first heat dissipation channel located at the bottom of the first temperature control component 110 and a heat dissipation fan (not shown in the figure) disposed in the first heat dissipation channel. Thus, when cooling is required, the heat of the first temperature control component 110 and the microfluidic chip 500 can be taken away more quickly through the first heat dissipation component 120, so as to facilitate rapid cooling, thereby facilitating more accurate and rapid realization of PCR reaction conditions, and facilitating accurate PCR reaction under specific temperature conditions.

[0075] In one embodiment, Figure 1 to Figure 3 As shown, the optical detection module 200 includes:

[0076] An optical detection component 210, wherein the optical detection component 210 is capable of detecting the PCR reaction result in the reaction area of ​​the microfluidic chip 500;

[0077] The optical detection moving component 220 can drive the optical detection component 210 to move, so as to detect the PCR reaction results in different microfluidic chips 500 and / or in different reaction chambers of the microfluidic chip 500.

[0078] Regarding the optical detection component, it can specifically be a lens unit, a filter, and a photosensitive chip (CMOS sensor) arranged in sequence from one side to the other, so as to take pictures of the reaction area of ​​the microfluidic chip (such as taking pictures of the fluorescence after the PCR reaction), so as to obtain the detection results based on the photographed results, including information such as the size of the droplets and the fluorescence intensity of the droplets.

[0079] Regarding the optical detection moving component 220, the present application has no specific limitation, as long as it can drive the optical detection component 210 to move. Figure 3 As shown, in this embodiment, the optical detection assembly is driven to move by a lead screw, specifically, the motor (first motor 221) drives the screw (first screw 222) of the lead screw to rotate through a belt, thereby driving the nut set on the screw to move, so as to drive the optical detection assembly 210 fixed to the nut to move. More specifically, in this embodiment, the setting direction of the screw (first screw 222) of the lead screw is perpendicular to the direction in which the chip moving module drives the microfluidic chip to move, so that each microfluidic chip 500 arranged in a row can be detected in turn.

[0080] In addition, the optical detection module may further include a second heat dissipation channel 230 for dissipating heat for the optical detection component 210 .

[0081] In one embodiment, Figure 1 to Figure 3 , Figure 5 As shown, the pressure plate module 300 also includes:

[0082] The first pressure plate moving component can drive the air supply component 310 to move to the upper part of the temperature control module 100 and move out from the upper part of the temperature control module 100. In the present embodiment, it specifically moves in the horizontal direction, and more specifically can move between the temperature control module 100 and the optical detection module 200.

[0083] Specifically, in this embodiment, the first pressing plate moving assembly includes:

[0084] A carrier frame 321, wherein the carrier frame 321 is capable of carrying the air supply assembly 310;

[0085] A first guide rail 322, wherein the first guide rail 322 is disposed on the rack (in this embodiment, there are specifically two first guide rails 322, which are disposed on both sides of the rack 900 respectively);

[0086] A slider 323, wherein the slider 323 is disposed at the bottom of the carrier 321;

[0087] The first moving unit 324 , wherein the moving end of the first moving unit 324 is directly or indirectly connected to the slider 323 , so as to drive the air supply assembly 310 disposed on the supporting frame 321 to slide along the first guide rail 322 through the slider 323 .

[0088] The present application has no specific restrictions on the first motion unit. Specifically, for example, a cylinder (those skilled in the art know that the push rod of the cylinder is its moving end, and the cylinder body of the cylinder is its fixed end), a lead screw (those skilled in the art know that the motor of the lead screw can be regarded as its fixed end, and the nut set on the screw can be regarded as its moving end). Figure 3 As shown, in this embodiment, a lead screw is used, specifically, a motor (second motor 325) is used to drive the screw (second screw 326) of the lead screw to rotate through a belt, thereby driving the nut set on the screw (second screw 326) to move, and the nut is fixed on the support frame 321, so as to realize that the air supply assembly 310 set on the support frame 321 is driven by the first motion unit 324 to slide along the first guide rail 322 through the slider 323.

[0089] In one embodiment, Figure 5 As shown, the pressure plate module 300 also includes:

[0090] The second pressure plate moving component, when the first pressure plate moving component drives the gas supply component 310 to move to the upper part of the temperature control module 100, the second pressure plate moving component can drive the gas supply component 310 to move to the gas supply position of the microfluidic chip 500 (that is, the position that can provide air pressure to the microfluidic chip 500) or move out of the gas supply position of the microfluidic chip 500 (in this embodiment, the second pressure plate moving component specifically drives the gas supply component 310 to move in the vertical direction (up and down direction) to move to or out of the gas supply position).

[0091] Wherein, the second pressing plate moving assembly comprises:

[0092] The second moving unit 331 , the moving end of the second moving unit 331 is directly or indirectly connected to the gas supply component 310 to drive the gas supply component to move to the gas supply position of the microfluidic chip or move out of the gas supply position of the microfluidic chip.

[0093] In order to facilitate the setting of the second pressure plate moving assembly, in this embodiment, the support frame 321 includes: a fixed plate 327, to which the fixed end of the second moving unit 331 is connected; and a guide column 328, which connects the slider and the fixed plate.

[0094] The second pressure plate moving assembly also includes: 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 to the bottom of the movable plate 332; the fixed end of the second motion unit is connected to the fixed plate 327; the moving end of the second motion unit 331 is connected to the movable plate (that is, in this embodiment, the moving end of the second motion unit is indirectly connected to the air supply assembly 310 through the movable plate 332).

[0095] Regarding the second motion unit 331, the present application has no specific restrictions, as long as it can drive the gas supply component to move to the gas supply position of the microfluidic chip or move out of the gas supply position of the microfluidic chip. Specific examples include a cylinder (the push rod of the cylinder is its moving end, and the cylinder body of the cylinder is its fixed end), a lead screw (the motor of the lead screw can be regarded as its fixed end, and the nut set on the screw can be regarded as its moving end). Figure 3 As shown, in this embodiment, a lead screw is used, specifically (two) motors (the third motor 333) arranged on the upper part of the fixed plate drive the vertically arranged screw (the third screw 334) to rotate, so as to drive the movable plate 332 to move up and down, thereby driving the air supply assembly 310 fixed at the bottom of the movable plate 332 to move to the air supply position of the microfluidic chip 500 or move out of the air supply position of the microfluidic chip 500.

[0096] In one embodiment, Figures 9 to 11 As shown, the pressure plate module 300 also includes:

[0097] The second heating component 340, when the second pressure plate moving component drives the gas supply component 310 to move to the gas supply position of the microfluidic chip 500, the second heating component 340 can heat the opposite side of the temperature control module 100 of the microfluidic chip 500 (in this embodiment, the upper surface of the microfluidic chip 500).

[0098] In the current PCR heating process (such as flat plate heating), the heating module is generally only located below the microfluidic chip, so the heat is conducted from bottom to top into the chip chamber or from top to bottom out of the chip chamber, and the heat will quickly lose from above the chip, making it unable to quickly reach or maintain the set temperature. In this embodiment, a second heating component is provided, which is located above the chip and contacts the chip during droplet generation and PCR reaction.

[0099] Specifically, the upper end of the second heating component 340 is connected with an elastic member 341 (a spring in this embodiment), and the elastic member 341 is directly or indirectly (in this embodiment, indirectly through the movable plate 332) connected to the moving end of the second moving unit 331 (that is, in this embodiment, the air supply component 310 and the elastic member 341 used to connect the second heating component 340 are both arranged on the movable plate 332); the air outlet of the air supply component 310 (the air outlet connecting the chip inlet and outlet) is located on both sides of the second heating component 340, that is, the second heating component 340 is located between the air outlets (the air outlet connecting the chip inlet and outlet). Therefore, the air outlets of the air supply components 310 located on both sides can output air pressure to the inlets and outlets on both sides of the microfluidic chip 500 or stop outputting air pressure, and the second heating component 340 located in the middle can heat the reaction area located in the middle of the microfluidic chip 500.

[0100] The second heating assembly 340 includes more than one (e.g., 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 zone of the microfluidic chip 500. Thus, each microfluidic chip 500 can be temperature-controlled independently to perform PCR reactions under different temperature conditions.

[0101] The present application has no specific limitation on the second temperature control unit 342, as long as it can heat the microfluidic chip. In this embodiment, the second temperature control unit 342 is an electric heater.

[0102] Furthermore, if Fig.10 , Fig.11 As shown, the second heating unit 342 is provided with a plurality of blind holes 343 facing the reaction area of ​​the microfluidic chip, thereby forming a heat preservation mechanism, which greatly reduces the conduction of heat such as heat convection, realizes the heat preservation of the chip surface, makes it closer to the temperature of the semiconductor cooler on the lower surface of the chip, ensures that the required temperature is maintained inside the chip, and realizes more precise control of the droplet generation temperature and PCR temperature.

[0103] In addition, the second temperature control unit 342 may also be a pressure plate with a heat-insulating material or a heat-insulating structure, which can quickly adjust the temperature in the chip chamber or maintain the set temperature by reducing the heat dissipation above the microfluidic chip.

[0104] In one embodiment, Figure 12 to Figure 14 As shown, the pressure plate module 300 is provided with relatively arranged limiting grooves 350. Specifically, in this embodiment, the limiting grooves 350 are respectively opened on two vertically arranged plates;

[0105] The pressing plate module 300 further includes:

[0106] The baffle 360 ​​includes:

[0107] Baffle body 361;

[0108] First protrusions 362, the first protrusions 362 are relatively arranged on both sides of the baffle body 361, the baffle body 361 is located at the inner sides of the relatively arranged limiting grooves 350 and the first protrusions 362 are respectively located in the limiting grooves 350;

[0109] A second protrusion 363, which is disposed at the bottom of the baffle body 361;

[0110] 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.

[0111] During optical detection, the excitation light needs to enter the chip from the bottom of the chip or from the top of the chip. If the excitation light enters the chip from the side of the chip, noise will be generated, affecting the scanning detection.

[0112] When using the digital PCR all-in-one machine of the present application, when the microfluidic chip is heated to perform a PCR reaction, as Fig.13 As shown, due to the support of the second protrusion 363 located below the baffle body 361 by the surface of the temperature control module, only the second protrusion 363 below the baffle contacts the surface of the temperature control module 100, thus ensuring that the contact area between the baffle and the temperature control module 100 is very small and limited to the second protrusion 363, thereby greatly reducing the heat loss through the baffle. When performing optical detection, such as Fig.14 As shown, the microfluidic chip 500 moves 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 effect of gravity, the baffle will fall a certain distance. At this time, the side of the chip is blocked by the baffle body 361 to prevent the excitation light from entering from the side of the chip, thereby ensuring the accuracy of optical detection.

[0113] In one embodiment, Fig.12 As shown, the pressure plate module 300 also includes an elastic component 370, which is directly or indirectly fixed to the bottom of the movable plate 332. When the air supply component 310 is located at the air supply position of the microfluidic chip 500, the bottom of the elastic component 370 is located at the top of the microfluidic chip 500 or the tray 600 for loading the microfluidic chip and is in a pressurized state.

[0114] During the PCR process, the microfluidic chip 500 is heated and subjected to pressure from the platen module 300 (such as the gas supply component 310 and the second heating unit 342). Therefore, it is easy to adhere to the bottom of the platen module 300 (such as the gas supply component 310 and the second heating unit 342). After the elastic component 370 of this embodiment is set, when the gas supply component 310 leaves the gas supply position of the microfluidic chip 500, the elastic component will push the microfluidic chip 500 away from the bottom of the gas supply component 310, thereby preventing the microfluidic chip 500 from adhering to the bottom of the platen module, so as to facilitate the digital PCR integrated machine of this application to automatically replace the microfluidic chip 500.

[0115] Specifically, in this embodiment, Fig.12 As shown, the elastic component 370 includes a plurality of elastic units, which include a compression spring and a movable column arranged in a vertical through hole, the movable column is provided with protrusions at the top and bottom, and the compression spring is arranged between the lower part of the through hole and the protrusion at the bottom of the movable column.

[0116] In one embodiment, the air supply component 310 is capable of moving between the carrying position of the temperature control module 100 and the detection position of the optical detection module 200 (such as achieved by the above-mentioned first pressure plate moving component and the second pressure plate moving component); the chip moving module 400 is directly or indirectly fixedly connected to the air supply component 310 (in this embodiment, the air supply component 310 and the chip moving module 400 are both fixed to the movable plate 332, thereby achieving indirect fixation).

[0117] In this embodiment, the chip moving module 400 is fixed to the gas supply component 310. When the chip moving module 400 fixes the microfluidic chip or the tray 600 for loading the microfluidic chip, when the gas supply component 310 moves between the bearing position of the temperature control module 100 and the optical detection module 200, the microfluidic chip 500 can be moved from the temperature control module 100 to the detection position of the optical detection module 200 through the chip moving module 400, so as to facilitate the optical detection module to detect the microfluidic chip. That is, in this embodiment, the chip moving module 400 drives the microfluidic chip 500 from the bearing position of the temperature control module to the detection position of the optical detection module through the gas supply component 310 moving between the temperature control module 100 and the optical detection module 200.

[0118] Of course, a chip moving module completely independent of the pressure plate module can also be provided, and after the reagents in the microfluidic chip 500 complete the PCR, the chip moving module independently drives the microfluidic chip 500 from the temperature control module 100 to the detection position of the optical detection module 200. Compared with this, the solution of this embodiment is conducive to the miniaturization of the device and the simplification of the structure.

[0119] There is no specific restriction on the way in which the chip moving module 400 fixes the microfluidic chip 500 or the tray 600 for loading the microfluidic chip, for example, it can be achieved by structures such as magnetic adsorption or clamping. In the present embodiment, the chip moving module 400 includes a chip bracket assembly, one end of which is directly or indirectly fixedly connected to the gas supply assembly 310 (in the present embodiment, it is arranged on the movable plate 332 and is indirectly fixedly connected to the gas supply assembly 310), and the other end extends to the lower part of the gas supply assembly 310, so that when the second pressure plate moving assembly drives the gas supply assembly to move out of the gas supply position of the microfluidic chip, the chip bracket 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.

[0120] In one implementation of the chip moving module 400 of this embodiment, as Figure 5 , Figure 6 As shown, the chip holder assembly includes: a chip holder body 410, one end of the chip holder body 410 is directly or indirectly fixedly connected to the gas supply assembly 310 (in the present embodiment, it is arranged on the movable plate 332 and is indirectly fixedly connected to the gas supply assembly 310); a third motion unit 420, which is arranged on the chip holder body; a clamp 430, the clamp 430 is arranged at the moving end of the third motion unit 420, and when the gas supply assembly is located at the gas supply position of the microfluidic chip, the moving end of the third motion unit can drive the clamp to clamp the tray 600 for loading the microfluidic chip. In this embodiment, grooves are provided on both sides of the tray 600. When the moving end of the third moving unit 420 drives the clamp 430 to clamp the microfluidic chip, the clamp 430 penetrates into the groove, so that when the second moving 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 carrying position of the temperature control module 100, and then, the first pressure plate moving component can drive the air supply component 310 and the microfluidic chip 500 to move toward the optical detection module together.

[0121] Regarding the third motion unit 420, the present application has no specific restrictions, as long as it can drive the clamping claw 430 to clamp or release the microfluidic chip 500 or the tray 600 for loading the microfluidic chip. Specifically, for example, a cylinder (the push rod of the cylinder is its moving end, and the cylinder body of the cylinder is its fixed end), a lead screw (the motor of the lead screw can be regarded as its fixed end, and the nut set on the screw can be regarded as its moving end). Figure 6As shown, in this embodiment, two relatively arranged lead screws respectively drive the relatively arranged clamping claws 430 to move toward or 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 platen moving assembly drives the air supply assembly to move out of the air supply position and the first platen moving assembly drives the air supply assembly to move to the optical detection module, the tray 600 can be lifted by the chip bracket unit 440, and at the same time, the tray can be tightly fixed by the chip moving module (the clamping claws 430 clamp 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 results.

[0122] In another implementation of the chip moving module 400 of this embodiment, Figure 7 , Figure 8 As shown, the chip bracket assembly includes: two chip bracket units 440 arranged in pairs, one end of the chip bracket unit 440 is directly or indirectly fixedly connected to the gas supply assembly 310 (in this embodiment, it is fixedly connected to the movable plate 332 and indirectly connected to the gas supply assembly 310), and the other end of the chip bracket unit 440 extends to the lower part of the gas supply assembly 310; a third protrusion 450 is arranged on the upper surface of the other end of the chip bracket unit 440. At this time, the third protrusion 450 corresponds to the through hole or the blind hole arranged on the bottom surface of the tray 600, 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 can drive the gas supply assembly to move to the optical detection module, the tray 600 can be lifted by the chip bracket unit 440, and at the same time, 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 blind hole arranged on the tray 600, so as to increase the reliability of the equipment operation and the accuracy of the detection result.

[0123] Those skilled in the art know that the vertical distance between the upper part of the third protrusion 450 and the lower part of the air supply assembly 310 should be greater than the thickness of the tray 600 used, so that the tray 600 can be accommodated between the two. In this application, "vertical distance" refers to the distance between the feet of perpendicular lines drawn through two points relative to a vertical line.

[0124] In addition, the digital PCR all-in-one machine of the present application may further be provided with a display 800, such as a touch display, to input instructions and output test results.

[0125] The digital PCR integrated machine of the present application may further include a control module 700 to control the above modules. Specifically, the control module 700 is electrically connected to each motor (such as a stepper motor), a heating element, a photosensitive chip and / or a display in the above modules for automatic control. Regarding the control module, a module controlled by a single-chip microcomputer can be used, or a PLC control module can be used. On the basis of the above technical solutions given in this application, those skilled in the art know how to realize the electrical connection and control of the control module 700 with each motor, heating unit, photosensitive chip and / or display according to the prior art, and this application will not repeat them here.

[0126] Although the embodiments of the present application are described above, the present application is not limited to the above specific embodiments and application fields, and the above specific embodiments are merely illustrative and instructive, rather than restrictive. A person of ordinary skill in the art can make many forms under the guidance of this specification and without departing from the scope of protection of the claims of the present application, all of which belong to the scope of protection claimed in the present application.

Claims

1. A digital PCR all-in-one machine, characterized in that: include: A temperature control module, wherein the bearing position of the temperature control module can bear a microfluidic chip or a tray for bearing the microfluidic chip, and the temperature control module can control the temperature of a reaction area of ​​the microfluidic chip located at the bearing position of the temperature control module to perform a PCR reaction; An optical detection module, which is disposed on one side of the temperature control module and is capable of detecting a PCR reaction result in a reaction area of ​​the microfluidic chip; A pressing plate module, the pressing plate module comprising an air supply component, the air supply component can be moved to the upper part of the temperature control module and moved out from the upper part of the temperature control module; when the air supply component moves to the upper part of the temperature control module, the air supply component can output air pressure to the inlet of the microfluidic chip located at the bearing position of the temperature control module or stop outputting air pressure; A chip moving module, wherein the chip moving module can move the microfluidic chip from the bearing position of the temperature control module to the 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, characterized in that: The temperature control module includes a first temperature control component, which can control the temperature of at least one surface of the microfluidic chip located at the bearing position of the temperature control module.

3. The digital PCR all-in-one machine according to claim 2, characterized in that: The first temperature control component includes more than one first temperature control unit, and each of the first temperature control units corresponds to a reaction area of ​​the microfluidic chip.

4. The digital PCR all-in-one machine according to claim 2, characterized in that: The temperature control module also includes a first heat dissipation component; The first heat dissipation component comprises: a first heat dissipation channel, the first heat dissipation channel is located at the bottom of the first temperature control component; and a heat dissipation fan, the heat dissipation fan is arranged in the first heat dissipation channel.

5. The digital PCR all-in-one machine according to claim 1, characterized in that: The optical detection module comprises: An optical detection component capable of detecting a PCR reaction result in a reaction area of ​​the microfluidic chip; The optical detection moving component can drive the optical detection component to move so as to detect the PCR reaction results in different microfluidic chips and / or different reaction areas of the microfluidic chip.

6. The digital PCR all-in-one machine according to claim 1, characterized in that: When the air supply component moves to the upper part of the temperature control module, the air supply component can also output air pressure to the outlet of the microfluidic chip located at the bearing position of the temperature control module or stop outputting air pressure.

7. The digital PCR all-in-one machine according to claim 1, characterized in that: The platen module also includes: The first pressing plate moving assembly can drive the air supply assembly to move to the upper part of the temperature control module and move out from the upper part of the temperature control module.

8. The digital PCR all-in-one machine according to claim 7, characterized in that: The first platen moving assembly comprises: A carrier frame, the carrier frame being capable of carrying the air supply assembly; a first guide rail, wherein the first guide rail is arranged on the frame; A slider, the slider being arranged at the bottom of the carrier; A first moving unit, wherein a moving end of the first moving unit is directly or indirectly connected to the slider to drive the air supply assembly disposed on the carrier to slide along the first guide rail through the slider.

9. The digital PCR all-in-one machine according to claim 8, characterized in that: The platen module also includes: The second pressure plate moving component can drive the gas supply component to move to the gas supply position of the microfluidic chip or move out of the gas supply position of the microfluidic chip when the first pressure plate moving component drives the gas supply component to move to the upper part of the temperature control module.

10. The digital PCR integrated machine according to claim 9, characterized in that: The second platen moving assembly includes: A second motion unit, wherein a motion end of the second motion unit is directly or indirectly connected to the gas supply component to drive the gas supply component to move to the gas supply position of the microfluidic chip or move out of the gas supply position of the microfluidic chip.

11. The digital PCR integrated machine according to claim 10, characterized in that: The carrier frame comprises: a fixing plate; and, A guide post connecting the slider and the fixing plate; The second platen moving assembly also includes: A movable plate, wherein a guide hole is arranged on the movable plate, wherein the guide hole is sleeved on the guide column, and the movable plate can slide along the guide column; the fixed end of the second motion unit is connected to the fixed plate; the moving end of the second motion unit is connected to the movable plate; and the air supply assembly is fixed to the bottom of the movable plate.

12. The digital PCR integrated machine according to claim 10, characterized in that: The pressure plate module also includes: The second heating component, when the second pressure plate moving component drives the gas supply component to move to the gas supply position of the microfluidic chip, the second heating component can heat the opposite side of the temperature control module of the microfluidic chip.

13. The digital PCR integrated machine according to claim 12, characterized in that: The upper end of the second heating assembly is connected to an elastic member, and the elastic member is directly or indirectly fixedly connected to the moving end of the second moving unit; The air outlets of the air supply component are located on both sides of the second heating component.

14. The digital PCR integrated machine according to claim 12, characterized in that: The second heating assembly includes more than one second heating unit; Each of the second heating units corresponds to a reaction area of ​​the microfluidic chip.

15. The digital PCR integrated machine according to claim 14, characterized in that: The second heating unit is provided with a plurality of blind holes facing the reaction area of ​​the microfluidic chip.

16. The digital PCR all-in-one machine according to claim 1, characterized in that: The pressure plate module is provided with two limiting grooves arranged opposite to each other; The pressure plate module further includes a baffle, and the baffle includes: A baffle body, the baffle body being located at the inner side of the relatively arranged limiting grooves; More than two first protrusions, wherein 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, wherein the more than two second protrusions are arranged at the bottom of the baffle body; 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.

17. The digital PCR integrated machine according to claim 11, characterized in that: The pressure plate module also includes: An elastic component is directly or indirectly fixed to the bottom of the movable plate. When the air supply component is located at the air supply position of the microfluidic chip, the bottom of the elastic component is located at the top of the microfluidic chip or a tray for loading the microfluidic chip and is in a compressed state.

18. The digital PCR all-in-one machine according to claim 9, characterized in that: The air supply assembly can move between the carrying 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 to the air supply assembly.

19. The digital PCR integrated machine according to claim 18, characterized in that: The chip moving module comprises: A chip holder assembly, one end of which is directly or indirectly fixedly connected to the air supply assembly, and the other end of which extends to the lower part of the air supply assembly, so that when the second pressure 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 for loading the microfluidic chip and make the microfluidic chip leave the carrying position of the temperature control module.

20. The digital PCR integrated machine according to claim 19, characterized in that: The chip carrier assembly at least comprises: Two chip bracket bodies are arranged in pairs, one end of each chip bracket body is directly or indirectly fixedly connected to the gas supply assembly; a third motion unit, the third motion unit being disposed on the chip bracket body; The clamp is arranged at the moving end of the third moving unit. When the gas supply component is located at the gas supply position of the microfluidic chip, the moving end of the third moving unit can drive the clamp to clamp the tray for loading the microfluidic chip.

21. The digital PCR integrated machine according to claim 19, characterized in that: The chip carrier assembly comprises: Two chip bracket units are arranged in pairs, one end of each chip bracket unit is directly or indirectly fixedly connected to the air supply assembly, and the other end of each chip bracket unit extends to the lower part of the air supply assembly.

22. The digital PCR integrated machine according to claim 21, characterized in that: The chip carrier assembly further comprises: A third protrusion is provided on the upper surface of the other end of the chip carrier unit.

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