Micro-droplet generation and amplification device and control method thereof

Abstract

The invention provides a micro-droplet generation and amplification device and a control method thereof.The device comprises an integration module and a rotating mechanism, the integration module comprises a first working platform and a second working platform, and the first working platform is provided with a chip containing groove; the second working platform has a working state close to the first working platform and a chip taking and placing state far away from the first working platform, and the second working platform is provided with a liquid drop generation module and a PCR amplification module; the liquid drop generation module is in pressing butt joint with a chip oil liquid hole and a chip gas liquid hole of the integrated micro-liquid drop chip so as to form micro-liquid drops in the integrated micro-liquid drop chip, and a reaction chamber of the integrated micro-liquid drop chip is located in a temperature control cavity of the PCR amplification module; the rotating mechanism is used for driving the two working platforms to synchronously turn over by 180 degrees when the second working platform is in the working state. According to the invention, the structure is more compact, the size of the instrument is smaller, the portability is higher, and the manufacturing cost is relatively reduced.

Description

Technical Field

[0001] The present invention belongs to the technical field of digital PCR instrument design, and in particular relates to a micro-droplet generation and amplification device and a control method thereof. Background Art

[0002] Digital PCR is the latest quantitative technology. It is based on the single-molecule PCR method to count nucleic acid quantification. It is an absolute quantitative method. It mainly adopts the microfluidics or droplet method in the current hot research field of analytical chemistry to disperse a large amount of diluted nucleic acid solution into the microreactor or droplet of the biochip. The number of nucleic acid templates in each reactor is less than or equal to 1. In this way, after the PCR cycle, the reactor with a nucleic acid molecule template will give a fluorescent signal, and the reactor without a template will not have a fluorescent signal. According to the relative proportion and the volume of the reactor, the number of template copies in the sample can be accurately calculated.

[0003] Existing digital PCR systems are often complex in structure, resulting in large size and high cost. Not only do they take up more valuable laboratory space, but the high cost of instruments has also affected the popularity of digital PCR technology to a certain extent. In addition, nucleic acid amplification is a key link in digital PCR technology, and its amplification efficiency directly affects the accuracy of the final test results, and the efficiency of nucleic acid amplification is mainly affected by the accuracy and consistency of temperature control. Since digital PCR technology requires microdroplet samples to be heated through microfluidic chips, the heat transfer efficiency is limited by the chip structure and material heat transfer performance. A certain constant temperature time is required to make the temperature between the microdroplets uniform, which places higher requirements on the temperature uniformity of the temperature control module.

[0004] The invention patent with patent application number 202111381197.4 discloses an integrated digital PCR instrument and its control method, which has a high degree of automation and integration, can improve the efficiency of detection and analysis operations, and reduce labor costs. The technical advantages, in this technical scheme, are dispatched by a manipulator (also known as a dispatching mechanism) to dispatch the chip in turn between the micro-droplet generation module, the flipping module, the PCR amplification module, the detection module and the waste box. Multiple chips work at the same time to form an assembly line working mode. The chip completes the four digital PCR links of droplet generation, chip flipping, droplet amplification, and droplet fluorescence detection in turn. In this technical scheme, since the manipulator needs to dispatch in multiple modules, the overall structure of the instrument is not compact enough, the structural design is relatively complex, the size is large, the portability is poor, and the manufacturing cost is relatively high. Summary of the invention

[0005] Therefore, the technical problem to be solved by the present invention is to provide a micro-droplet generation and amplification device and a control method thereof, which effectively overcomes the shortcomings of the digital PCR instrument in the related technology in the prior art, such as complex structural design, large instrument size, lack of compactness, poor portability and high manufacturing cost.

[0006] In order to solve the above problems, the present invention provides a micro-droplet generation and amplification device, including an integrated module and a rotating mechanism, the integrated module including a first working platform and a second working platform, the first working platform having a plurality of chip placement grooves for positioning and placing an integrated micro-droplet chip, the second working platform being arranged in parallel with the first working platform and having a working state close to the first working platform and a chip placement state away from the first working platform, the second working platform having a droplet generation module and a PCR amplification module, when the second working platform is in the working state, the droplet generation module is respectively pressed and docked with the chip oil-liquid hole and the chip gas-liquid hole of the integrated micro-droplet chip so as to form micro-droplets in the integrated micro-droplet chip, and the reaction chamber of the integrated micro-droplet chip is in the temperature control cavity of the PCR amplification module, and the rotating mechanism is used to drive the first working platform and the second working platform to synchronously flip 180° when the second working platform is in the working state.

[0007] In some embodiments, the integrated module also includes vertical plates arranged at intervals on the left and right, the length ends of the first working platform are pivotally mounted on the top positions of the two vertical plates, and the bottom ends of the two vertical plates are connected as a whole through a connecting plate, the rotating mechanism includes a rotary motor and a transmission assembly, the rotary motor is fixedly connected to one of the vertical plates, and the rotary motor is driven and connected to one end of the first working platform via the transmission assembly to drive the first working platform to rotate a preset angle; and / or, the four corner areas corresponding to each other of the first working platform and the second working platform are each vertically provided with a guide slide bar, the outer periphery of the guide slide bar is sleeved with a first return spring, the first return spring is clamped between the first working platform and the second working platform, and also includes a downward pressing structure, the downward pressing structure can overcome the elastic force of the first return spring under the action of the downward pressure so that the second working platform switches from the chip placement state to the working state.

[0008] In some embodiments, the downward pressure structure includes a pressure rod and a roller hinged at both ends of the length of the second work platform. A traction rope is connected between the pressure rod and the first work platform. The traction rope forms a wrap angle on the roller. When the pressure rod is pressed toward one side of the first work platform, the traction rope can pull the first work platform and the second work platform towards each other and put them into the working state.

[0009] In some embodiments, when the pressure rod is pressed down to the lowest position toward the first working platform, the connection point between the traction rope and the second working platform is located on the side of the center point of the roller close to the first working platform.

[0010] In some embodiments, the droplet generation module includes a generation block, the length extension direction of the generation block is parallel to the width direction of the second working platform, and the two ends of the length of the generation block are connected to the second working platform via connecting ears, and the generation first hole and the generation second hole are formed on the generation block. The droplet generation module also includes a generation cover plate, and the generation cover plate has a covering position for sealing the top surface of the chip oil-liquid hole and the chip gas-liquid hole when generating micro-droplets and realizing the pressure-connected connection between the air path on the generation cover plate and the chip gas-liquid hole.

[0011] In some embodiments, the generated cover plate and the generated pressure block are hinged by a rotating shaft, and the generated pressure block is provided with an electromagnet. When the electromagnet is energized, the generated cover plate is attracted and rotates around the rotating shaft to be in the covered position; and / or, both ends of the length of the generated pressure block are vertically slidably connected to the second working platform via the connecting ears, and a guide rod is provided between the top surface of the generated pressure block and the bottom surface of the second working platform, and a third return spring is mounted on the outer surface of the guide rod, and the third return spring is clamped between the bottom surface of the second working platform and the top surface of the generated pressure block.

[0012] In some embodiments, there are two electromagnets, which are respectively located at the two ends of the length of the generated pressure block, and the two ends of the length of the generated cover plate respectively have suction blocks corresponding to the positions of the electromagnets; and / or, a second reset spring is mounted on the rotating shaft, which is used to switch the generated cover plate from the closed position to the open position when the electromagnet is powered off; and / or, a hand-holding protrusion is provided on the edge of one side of the generated cover plate away from the rotating shaft; and / or, a cover plate sealing gasket arranged around each of the generated first holes and generated second holes is provided on the end surface of the generated cover plate facing the generated pressure block.

[0013] In some embodiments, the PCR amplification module includes a semiconductor heater, a first end of the semiconductor heater is connected to a heat conductive block, the temperature control cavity is formed on the end surface of the heat conductive block facing away from the semiconductor heater, and a second end of the semiconductor heater is connected to a heat dissipation structure.

[0014] In some embodiments, the heat dissipation structure includes a first heat dissipation block contacting and connected to the second end and a second heat dissipation block contacting and connected to the first heat dissipation block, the heat dissipation area of ​​the second heat dissipation block is greater than the heat dissipation area of ​​the first heat dissipation block, the heat dissipation structure also includes a third heat dissipation block and a heat pipe, the first end of the heat pipe is between the first heat dissipation block and the second heat dissipation block, and the second end of the heat pipe is connected to the third heat dissipation block for heat exchange.

[0015] The present invention also provides a control method for the above-mentioned micro-droplet generation and amplification device, comprising the following steps: Placing the integrated micro-droplet chip containing the sample liquid into the chip placement groove; Controlling the second operation platform to switch from a chip pick-and-place state to a working state; Controlling the droplet generation module to operate so as to generate microdroplets in the integrated microdroplet chip, and storing the microdroplets in a reaction chamber of the integrated microdroplet chip; After the micro-droplets are generated, the first working platform and the second working platform are controlled to synchronously flip 180°, and the PCR amplification module is controlled to operate to amplify the micro-droplets in the reaction chamber. During the operation of the PCR amplification module, the first working platform and the second working platform are controlled to synchronously swing back and forth.

[0016] The present invention provides a micro-droplet generation and amplification device and a control method thereof, which integrates a droplet generation module and a PCR amplification module on a second working platform. The second working platform has a working state and a chip placement state and can switch between the two states. The first working platform and the second working platform can be synchronously flipped by the rotation drive of a rotating mechanism. There is no need to use a scheduling mechanism in the prior art to switch or flip the integrated micro-droplet chip between different working modules, so that the digital PCR instrument has a more compact structure and a smaller instrument size, thereby having stronger portability and relatively lower manufacturing costs. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is a schematic diagram of the structure of a compact digital PCR instrument according to an embodiment of the present invention (the overall structure omits the outer shell and other parts); Figure 2 for Figure 1 Schematic diagram of the three-dimensional structure of the integrated micro-droplet chip; Figure 3 for Figure 1 Schematic diagram of the three-dimensional structure of the integrated module (including the rotating mechanism); Figure 4 yes Figure 3 A partial enlarged view of the middle A; Figure 5 for Figure 1 Schematic diagram of the three-dimensional structure of the droplet generation module; Figure 6 It is a schematic diagram of the three-dimensional structure of the PCR amplification module; Figure 7 It is a schematic diagram of the three-dimensional structure after the flip mechanism is assembled; Figure 8 It is a schematic diagram of the three-dimensional structure of the fluorescence detection module and the detection crimping module assembled in the instrument; Fig. 9 for Figure 8 A schematic diagram of the three-dimensional structure of the detection crimping module; Fig.10 for Figure 8 Schematic diagram of the three-dimensional structure of the fluorescence detection module; Fig.11 It is a schematic diagram of the three-dimensional structure of the instrument before the micro-droplet is generated; Fig.12 It is a schematic diagram of the three-dimensional structure of the instrument in the state of micro-droplet generation; Fig.13 This is a schematic diagram of the three-dimensional structure of the instrument in the PCR amplification state when flipped 180°; Fig.14 Schematic diagram of the local three-dimensional structure under fluorescence detection state.

[0018] The reference numerals are: 1. First working platform; 11. Vertical plate; 12. Connecting plate; 121. Guide slide bar; 122. First reset spring; 123. Pressure bar; 124. Roller; 125. Traction rope; 126. Linear bearing; 13. Chip placement groove; 131. Guide spring; 2. Second working platform; 3. Droplet generation module; 31. Generate first hole; 32. Generate second hole; 33. Generate pressure block; 34. Generate cover plate; 35. Electromagnet; 36. Rotating shaft; 37. Second reset spring; 38. Guide bar; 39. Third reset spring; 310. Cover plate sealing pad; 4. PCR amplification module; 41. Semiconductor heating element; 42. Heat insulation board; 43. Heat conduction block; 44. First heat dissipation block; 47. The third heat sink; 48. The second heat sink; 49. The heat pipe; 5. The rotating mechanism; 51. The rotary motor; 52. The transmission assembly; 6. The fluorescence detection module; 61. The objective lens; 62. The optical path assembly; 63. The lifting motor; 64. The translation motor; 7. The detection crimping module; 70. The detection oil circuit; 71. The detection crimping block; 72. The moving block; 73. The first crimping motor; 74. The second crimping motor; 75. The first sealing gasket; 76. The air cavity; 77. The light source; 100. The integrated micro-droplet chip; 101. The chip oil-liquid hole; 102. The chip air-liquid hole; 103. The reaction chamber; 104. The fluorescence detection area; 105. The sample loading chamber; 201. The device bottom plate; 2021. The longitudinal guide rail; 2022. The longitudinal motor. DETAILED DESCRIPTION

[0019] See also Figures 1 to 14As shown, according to an embodiment of the present invention, a compact digital PCR instrument is provided, including a micro-droplet generation and amplification device (not labeled in the figure), the micro-droplet generation and amplification device includes an integrated module (not labeled in the figure) and a rotating mechanism 5 (not labeled in the figure), the integrated module includes a first working platform 1 and a second working platform 2, the first working platform 1 has a plurality of chip placement grooves 13 for positioning and placing an integrated micro-droplet chip 100, it can be understood that each chip placement groove 13 is used to position and place an integrated micro-droplet chip 100, the second working platform 2 is arranged in parallel with the first working platform 1 and has a working state close to the first working platform 1 and a chip placement state away from the first working platform 1, the second working platform 2 has a droplet generation module 3 and a PCR amplification module 4, when the second working platform 2 is in the working state, the droplet generation module 3 and the PCR amplification module 4 are provided on the second working platform 2. The droplet generation module 3 is pressed and docked with the chip oil-liquid hole 101 and the chip gas-liquid hole 102 of the integrated micro-droplet chip 100 respectively so as to form micro-droplets in the integrated micro-droplet chip 100. The formed micro-droplets will be stored in the reaction chamber 103 of the integrated micro-droplet chip 100, and the reaction chamber 103 of the integrated micro-droplet chip 100 is in the temperature control cavity (not shown and not labeled) of the PCR amplification module 4. The rotating mechanism 5 is used to drive the first working platform 1 and the second working platform 2 to synchronously flip 180° when the second working platform 2 is in the working state, so that the reaction chamber 103 is turned upside down, at this time, it is ensured that the micro-droplets stored in the reaction chamber 103 can be at the bottom of the reaction chamber 103 under the action of their own weight, thereby ensuring that the temperature control cavity of the PCR amplification module 4 can more efficiently control the temperature of the micro-droplets (that is, the reaction liquid).

[0020] In this technical solution, the droplet generation module 3 and the PCR amplification module 4 are integrated on the second working platform 2. At the same time, the second working platform 2 has a working state and a chip placement state and can switch between the two states. The rotation drive of the rotating mechanism 5 realizes the synchronous flipping of the first working platform 1 and the second working platform 2. There is no need to use the scheduling mechanism in the prior art to switch or flip the integrated micro-droplet chip 100 between different working modules, so that the structure of the digital PCR instrument is more compact and the instrument size is smaller, so it has stronger portability, and the manufacturing cost is relatively reduced.

[0021] See Figure 7As shown, the shape of the aforementioned chip placement groove 13 matches the bottom surface shape of the integrated micro-droplet chip 100. As a preferred embodiment, a corresponding guide spring 131 is provided on the side of the aforementioned chip placement groove 13 away from the operating side of the operator, so that the position of the chip can be more reliable when the chip is placed, and when the chip is taken out, force can be applied to the end of the chip to facilitate the chip to be ejected from the chip placement groove 13 and thus facilitate the removal of the chip.

[0022] In some embodiments, the integrated module also includes a vertical plate 11 spaced apart on the left and right, and the vertical plates 11 have two and are placed vertically spaced apart. The first working platform 1 is pivoted at both ends of its length (for example, connected by a rolling shaft) and mounted on the top positions of the two vertical plates 11. The bottom ends of the two vertical plates 11 are connected as a whole through a connecting plate 12 to improve the structural reliability between the two vertical plates 11. The rotating mechanism 5 includes a rotary motor 51 and a transmission assembly 52. ​​The rotary motor 51 is fixedly connected to one of the vertical plates 11. The rotary motor 51 is driven and connected to one end of the first working platform 1 via the transmission assembly 52 to drive the first working platform 1 to rotate by a preset angle. In a specific embodiment, the rotary motor 51 and the transmission assembly 52 can be implemented using a rotary drive assembly available on the market.

[0023] In this technical solution, the rotating mechanism 5 can drive the first working platform 1 to flip up and down by 180°, so that the top surface of the integrated micro-droplet chip 100 faces upward to generate micro-droplets, and after the micro-droplets are generated, the first working platform 1 and the second working platform 2 are rotated to flip up and down synchronously, so that the top surface of the integrated micro-droplet chip 100 faces downward to achieve that the micro-droplets in its reaction chamber 103 can be at the bottom of the reaction chamber 103 under the action of their own weight, thereby ensuring that the PCR amplification module 4 can reliably control the temperature of the micro-droplets in the reaction chamber 103. At the same time, the rotating mechanism 5 can also drive the first working platform 1 to swing back and forth during the amplification process, that is, rotate the aforementioned preset angle (generally ±10°), so that the uniform process of the temperature field accelerated by fluid convection can be improved, the heat transfer efficiency and temperature uniformity in the micro-droplet sample can be improved, and the temperature uniformity requirement of the temperature control module itself can be reduced, thereby improving the efficiency of the amplification link.

[0024] In some embodiments, see Figure 8As shown, the compact digital PCR instrument also includes a device base plate 201 and a longitudinal movement component (not marked in the figure), the longitudinal movement component includes a longitudinal movement guide rail 2021 and a longitudinal movement motor 2022, the bottom end of the integrated module is slidably connected to the device base plate 201 via the longitudinal movement guide rail 2021, and the longitudinal movement motor 2022 can drive the integrated module as a whole to move back and forth in a straight line along the guiding direction of the longitudinal movement guide rail 2021, and the guiding direction of the longitudinal movement guide rail 2021 is a horizontal direction perpendicular to the length direction of the first working platform 1.

[0025] In this technical solution, the longitudinal movement component can drive the integrated module to adjust its position in the horizontal longitudinal direction of the instrument, so that the integrated module can adjust its corresponding position with the corresponding processing module (such as the subsequent detection crimping module 7 and the fluorescence detection module 6) in different process flows, simplifying the structural design of the processing module. For example, there is no need for longitudinal displacement control of the processing module, that is, the control is simplified.

[0026] See also Figure 8 As shown, in some embodiments, the compact digital PCR instrument further includes a fluorescence detection module 6 on the first side of the integrated module and a detection crimping module 7 on the second side of the integrated module, wherein the first side and the second side are respectively one of the top side and the bottom side of the first working platform 1. In a specific embodiment, Figure 8 The orientation shown is for reference. The first side is the upper side of the integrated module, and the second side is the lower side of the integrated module. The fluorescence detection module 6 can be controlled to rise and fall to move away from or approach the first working platform 1 to achieve the position correspondence with the fluorescence detection area 104 of the integrated micro-droplet chip 100 and perform fluorescence detection. The detection crimping module 7 can be controlled to rise and fall to move closer to or away from the second working platform 2 to achieve compression docking with the first generation hole 31 and the second generation hole 32 in the droplet generation module 3, so that the detection oil in the detection oil circuit 70 (specifically, it can be understood as a detection oil bottle) can enter the reaction chamber 103 to float the micro-droplets after the amplification reaction to the fluorescence detection area 104 and enter the sample loading chamber 105 of the integrated micro-droplet chip 100. The fluorescence detection module 6 and the detection crimping module 7 can be controlled to move to correspond to the position of each integrated micro-droplet chip 100.

[0027] In this technical solution, by respectively arranging the fluorescence detection module 6 and the detection crimping module 7 on the upper and lower sides of the integrated module, the PCR instrument of the present invention can integrate the fluorescence detection function of the micro-droplets to achieve functional integration of the instrument.

[0028] See Fig. 9As shown, in some embodiments, the detection crimping module 7 includes an air cavity 76 and a crimping assembly (not labeled in the figure) connected to the air cavity 76, and the air cavity 76 is used to provide negative pressure for the droplet generation module 3 and provide positive pressure for the detection crimping module 7, wherein during the operation of the droplet generation module 3, negative pressure is provided for the micro-droplet generation process, and during the operation of the fluorescence detection module 6, positive pressure is provided for the supply of detection oil.

[0029] In this technical solution, the air cavity 76 can serve as a pressure vessel of the instrument and also as an assembly carrier of the crimping assembly, which can further simplify the structural design of the instrument and improve the compactness of the structure of the instrument.

[0030] Continue to see Fig. 9 As shown, in some embodiments, the crimping assembly includes a detection crimping block 71 and a moving block 72, the detection crimping block 71 is slidably connected to the moving block 72 and can be driven to rise and fall by the first crimping motor 73, the moving block 72 is slidably connected to the air cavity 76 and can be driven to move laterally along the length direction of the first working platform 1 by the second crimping motor 74, so as to realize the sequential docking of the crimping assembly to each of the plurality of integrated micro-droplet chips 100. It can be understood that the crimping assembly is also provided with corresponding guide rails to guide the smooth lifting and lateral movement of the detection crimping block 71; in a preferred embodiment, the moving block 72 is also provided with a light source 77 to provide a light field for the camera of the fluorescence detection module 6, and the light source 77 is arranged on the moving block 72, so that the relative position of the light source 77 and the detection crimping block 71 is stable, and the two move synchronously, and the control is simplified.

[0031] In some embodiments, the detection crimping block 71 is formed with a first detection through hole and a second detection through hole corresponding to the chip oil-liquid hole 101 and the chip gas-liquid hole 102 respectively, and the detection oil enters the reaction chamber 103 via the first detection through hole and the second detection through hole. The detection crimping block 71 has a first sealing gasket 75 for forming a seal at the crimping position when the detection crimping block 71 is crimped with the droplet generation module 3.

[0032] See also Figure 3 and Figure 4As shown, in some embodiments, guide slide bars 121 are vertically arranged at the four corresponding corner areas of the first working platform 1 and the second working platform 2. In a specific embodiment, a sliding connection is formed between the guide slide bar 121 and the first working platform 1 and / or the second working platform 2 through a linear bearing 126. A first return spring 122 is sleeved on the outer periphery of the guide slide bar 121. The first return spring 122 is clamped between the first working platform 1 and the second working platform 2 so as to apply a restoring elastic force to separate the first working platform 1 and the second working platform 2. The guide slide bar 121 also includes a downward pressing structure (not marked in the figure). The downward pressing structure can overcome the elastic force of the first return spring 122 under the action of the downward pressure so that the second working platform 2 is switched from the chip placement state to the working state. The aforementioned downward pressure can be specifically achieved by manual downward pressure by an operator.

[0033] In this technical solution, force is applied to the downward pressing structure to overcome the elastic force of the first return spring 122 to achieve the approach of the first working platform 1 and the second working platform 2, thereby achieving the crimping of the droplet generation module 3 and the integrated micro-droplet chip 100. When the downward pressing structure is subjected to reverse force, that is, an upward pulling force is applied, the first return spring 122 can assist the second working platform 2 to switch from the working state to the chip placement state, and prevent the downward pressing structure from falling under the action of its own weight, thereby facilitating the placement of the chip.

[0034] See Figure 4 As shown, in some embodiments, the pressing structure includes a pressure rod 123 and a roller 124 hinged at both ends of the length of the second working platform 2, and a traction rope 125 (such as a steel wire) is connected between the pressure rod 123 and the first working platform 1, and the traction rope 125 forms a wrap angle on the roller 124 (that is, the traction rope 125 partially wraps the outer circumferential surface of the roller 124). When the pressure rod 123 is pressed toward the side of the first working platform 1, the traction rope 125 can pull the first working platform 1 and the second working platform 2 towards each other and be in the working state. Preferably, when the pressure rod 123 is pressed toward the side of the first working platform 1 to the lowest position, the connection point of the traction rope 125 and the second working platform 2 is on the side of the center point of the roller 124 close to the first working platform 1.

[0035] In this technical solution, the state switching of the second working platform 2 can be driven when the position of the pressure rod 123 changes through the directional guiding function of the roller 124 and the pulling connection function of the rope. The structure is simple, especially after the aforementioned pressure rod 123 is pressed down, because the connection point between the traction rope 125 and the pressure rod 123 is below the center point of the roller 124, the self-locking of the structure is achieved, and there is no need to set up a corresponding locking device separately, which further simplifies the structural design.

[0036] See Figure 5 As shown, the droplet generation module 3 includes a generation block 33, the length extension direction of the generation block 33 is parallel to the width direction of the second working platform 2, and the two ends of the length of the generation block 33 are connected to the second working platform 2 via connecting ears (not marked in the figure), the first generation hole 31 and the second generation hole 32 are both formed on the generation block 33, and the droplet generation module 3 also includes a generation cover plate 34, and the generation cover plate 34 has a covering position for sealing the top surface of the chip oil-liquid hole 101 and the chip gas-liquid hole 102 through the cover plate sealing gasket 310 when generating micro-droplets and realizing the pressure-connected connection between the air path (not marked in the figure) on the generation cover plate 34 and the chip gas-liquid hole 102.

[0037] In this technical solution, both ends of the generated pressing block 33 are connected to the second working platform 2 via connecting lifting ears, and the structure is reliable and simple.

[0038] In some embodiments, the generation cover plate 34 is hinged to the generation pressing block 33 via a rotating shaft 36, and the generation pressing block 33 has an electromagnet 35. When the electromagnet 35 is energized, the generation cover plate 34 is attracted and rotated around the rotating shaft 36 to be in the covering position. This technical solution realizes the attraction of the generation cover plate 34 through the electromagnet 35, and the structure and control are relatively simple.

[0039] In some embodiments, there are two electromagnets 35, which are respectively located at two ends of the length of the generated pressing block 33, and the two ends of the length of the generated cover plate 34 are respectively provided with a suction block corresponding to the position of each electromagnet 35. The two electromagnets 35 are respectively located at two ends of the length of the generated pressing block 33 to ensure the smooth closure of the generated cover plate 34 and to ensure the reliable sealing of the generated cover plate 34.

[0040] In some embodiments, the rotating shaft 36 is provided with a second return spring 37, which is used to switch the generating cover plate 34 from the closing position to the opening position when the electromagnet 35 is powered off. In this technical solution, the second return spring 37 can be provided to open the generating cover plate 34 by its own elastic force when the electromagnet 35 is powered off. The second return spring 37 can be a torsion spring.

[0041] In some embodiments, a hand-held protrusion (not shown) is provided on one side edge of the generation cover plate 34 away from the rotating shaft 36, so that the operator can apply force to the hand-held protrusion to assist in opening or closing the generation cover plate 34. A cover plate sealing gasket 310 is provided on one side end surface of the generation cover plate 34 facing the generation pressing block 33, which is arranged around each of the generation first hole 31 and the generation second hole 32.

[0042] In some embodiments, the two ends of the length of the generating block 33 can be vertically slidably connected to the second working platform 2 via the connecting ears, and a guide rod 38 is provided between the top surface of the generating block 33 and the bottom surface of the second working platform 2. The third return spring 39 is sheathed on the guide rod 38, and the third return spring 39 is clamped between the bottom surface of the second working platform 2 and the top surface of the generating block 33. In this technical solution, while the two ends of the generating block 33 can be vertically slidably connected to the second working platform 2 via the connecting ears, the clamping of the third return spring 39 between the second working platform 2 and the generating block 33 can buffer and adjust the horizontal state of the generating block 33, thereby ensuring the crimping sealing of the generating block 33 to the integrated micro-droplet chip 100.

[0043] See Figure 6 As shown, in some embodiments, the PCR amplification module 4 includes a semiconductor heating element 41, a first end of the semiconductor heating element 41 is connected to a heat conductive block 43, the temperature control cavity is formed on the end surface of the heat conductive block 43 facing away from the semiconductor heating element 41, and the second end of the semiconductor heating element 41 is connected to a heat dissipation structure (not marked in the figure), and the aforementioned heat conductive block 43 can evenly transfer the heat of the semiconductor heating element 41 to the temperature control cavity, thereby being able to uniformly control the temperature of the micro-droplets in the reaction chamber 103. The aforementioned semiconductor heating element 41 specifically adopts a Peltier, and according to actual needs, a plurality of Peltiers can be used.

[0044] In some embodiments, the heat dissipation structure includes a first heat dissipation block 44 connected in contact with the second end and a second heat dissipation block 48 connected in contact with the first heat dissipation block 44, and the heat dissipation area of ​​the second heat dissipation block 48 is larger than the heat dissipation area of ​​the first heat dissipation block 44, so as to achieve rapid dissipation of heat from the first end of the semiconductor heating element 41.

[0045] In some embodiments, the heat dissipation structure further includes a third heat dissipation block 47 and a heat pipe 49. The heat pipe 49 may be a heat pipe, which contains a heat-conducting medium and has a high heat transfer efficiency. The first end of the heat pipe 49 is located between the first heat dissipation block 44 and the second heat dissipation block 48, and the second end of the heat pipe 49 is connected to the third heat dissipation block 47 for heat exchange. In this technical solution, the heat at the first heat dissipation block 44 and the second heat dissipation block 48 can be quickly transferred and conducted to the third heat dissipation block 47 through the heat pipe 49 for dissipation. The length of the heat pipe 49 can be adjusted to achieve flexible selection of the location of the third heat dissipation block 47, thereby greatly increasing the heat dissipation area and achieving the purpose of rapid heat dissipation.

[0046] In some embodiments, a heat insulation plate 42 is arranged in a circle at the connection position between the first heat sink 44 and the semiconductor heating element 41 to prevent short-circuiting of cold and hot water and reduce energy consumption; and / or, a temperature probe (not shown in the figure) and a temperature fuse (not shown in the figure) are arranged in the heat conductive block 43, wherein the temperature probe can detect the real-time temperature at the heat conductive block 43, and the temperature fuse can disconnect the power supply of the semiconductor heating element 41 when the real-time temperature of the heat conductive block 43 exceeds a preset value, thereby achieving the purpose of constant temperature control.

[0047] In some embodiments, the chip placement grooves 13 are arranged in sequence and at intervals along the length direction of the first working platform 1, and the droplet generation modules 3 and the PCR amplification modules 4 are arranged one-to-one corresponding to the chip placement grooves 13, thereby improving the processing throughput of the instrument.

[0048] According to an embodiment of the present invention, a control method for the above-mentioned compact digital PCR instrument is also provided, comprising the following steps: Place the integrated micro-droplet chip 100 loaded with the sample liquid into the chip placement groove 13; Controlling the second working platform 2 to switch from a chip picking and placing state to a working state; Controlling the droplet generation module 3 to operate so as to generate micro-droplets in the integrated micro-droplet chip 100, and storing the micro-droplets in the reaction chamber 103 of the integrated micro-droplet chip 100; After the micro-droplets are generated, the first working platform 1 and the second working platform 2 are controlled to synchronously flip 180 degrees, and the PCR amplification module 4 is controlled to operate to amplify the micro-droplets in the reaction chamber 103; After amplification is completed, the detection crimping module 7 and the fluorescence detection module 6 are controlled to move toward the integrated micro-droplet chip 100. After the detection crimping module 7 is docked with the first generation hole 31 and the second generation hole 32 of the droplet generation module 3, the detection crimping module 7 is controlled to operate to drive the amplified micro-droplets from the reaction chamber 103 through the fluorescence detection area 104 of the integrated micro-droplet chip 100 into the sample loading cavity 105 of the integrated micro-droplet chip 100, and the fluorescence detection module 6 is controlled to perform fluorescence detection in the fluorescence detection area 104.

[0049] This technical solution integrates the droplet generation module 3 and the PCR amplification module 4 on the second working platform 2. At the same time, the second working platform 2 has a working state and a chip placement state and can switch between the two states. The first working platform 1 and the second working platform 2 are synchronously flipped by the rotation drive of the rotating mechanism 5. There is no need to use the scheduling mechanism in the prior art to switch or flip the integrated micro-droplet chip 100 between different working modules, so that the structure of the digital PCR instrument is more compact and the instrument volume is smaller, so it has stronger portability, and the manufacturing cost is relatively reduced. In some embodiments, the control method also includes: during the operation of the PCR amplification module 4, controlling the first working platform 1 and the second working platform 2 to swing back and forth synchronously, which can improve the heat transfer efficiency and temperature uniformity in the micro-droplet sample through the uniform process of the temperature field accelerated by fluid convection, and also reduce the temperature uniformity requirements of the temperature control module itself, thereby improving the efficiency of the amplification link; and / or, after the fluorescence detection is completed, controlling the first working platform 1 and the second working platform 2 to rotate 180° in the opposite direction synchronously, and then controlling the second working platform 2 to switch from the working state to the chip placement state.

[0050] It is easy for those skilled in the art to understand that, under the premise of no conflict, the above-mentioned advantageous methods can be freely combined and superimposed.

[0051] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention shall be included in the protection scope of the present invention. The above are only preferred embodiments of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and variations can be made without departing from the technical principles of the present invention, and these improvements and variations should also be regarded as the protection scope of the present invention.

Claims

1. A micro-droplet generation and amplification device, characterized in that: The invention comprises an integrated module and a rotating mechanism (5), wherein the integrated module comprises a first working platform (1) and a second working platform (2), wherein the first working platform (1) has a plurality of chip placement grooves (13) for positioning and placing an integrated micro-droplet chip (100), the second working platform (2) is arranged in parallel with the first working platform (1) and has a working state close to the first working platform (1) and a chip taking and placing state far from the first working platform (1), the second working platform (2) has a droplet generation module (3) and a PCR amplification module (4), and when the second working platform ( 2) When in the working state, the droplet generation module (3) is pressed and docked with the chip oil-liquid hole (101) and the chip gas-liquid hole (102) of the integrated micro-droplet chip (100) so as to form micro-droplets in the integrated micro-droplet chip (100), and the reaction chamber (103) of the integrated micro-droplet chip (100) is located in the temperature control chamber of the PCR amplification module (4), and the rotating mechanism (5) is used to drive the first working platform (1) and the second working platform (2) to synchronously flip 180 degrees when the second working platform (2) is in the working state.

2. The micro-droplet generation and amplification device according to claim 1, characterized in that: The integrated module further comprises vertical plates (11) arranged at intervals on the left and right sides, the first working platform (1) is pivotally mounted at both ends of its length at the top positions of the two vertical plates (11), and the bottom positions of the two vertical plates (11) are connected as a whole via a connecting plate (12), the rotating mechanism (5) comprises a rotary motor (51) and a transmission assembly (52), the rotary motor (51) is fixedly connected to one of the vertical plates (11), and the rotary motor (51) is drivingly connected to one end of the first working platform (1) via the transmission assembly (52) to drive the first working platform (1) to rotate. A preset angle; and / or, the four corresponding corner areas of the first working platform (1) and the second working platform (2) are each vertically provided with a guide slide bar (121), the outer periphery of the guide slide bar (121) is sleeved with a first return spring (122), the first return spring (122) is clamped between the first working platform (1) and the second working platform (2), and also includes a downward pressing structure, the downward pressing structure can overcome the elastic force of the first return spring (122) under the action of the downward pressure so that the second working platform (2) switches from the chip placement state to the working state.

3. The micro-droplet generation and amplification device according to claim 2, characterized in that: The pressing structure comprises a pressure rod (123) and a roller (124) hinged at both ends of the length of the second working platform (2); a traction rope (125) is connected between the pressure rod (123) and the first working platform (1); the traction rope (125) forms a wrap angle on the roller (124); when the pressure rod (123) is pressed down toward one side of the first working platform (1), the traction rope (125) can pull the first working platform (1) and the second working platform (2) toward each other and put them into the working state.

4. The micro-droplet generation and amplification device according to claim 3, characterized in that: When the pressure rod (123) is pressed down to the lowest position toward the first working platform (1), the connection point between the traction rope (125) and the second working platform (2) is located on the side of the center point of the roller (124) close to the first working platform (1).

5. The micro-droplet generation and amplification device according to claim 1, characterized in that: The droplet generation module (3) comprises a generation block (33), the length extension direction of the generation block (33) is parallel to the width direction of the second working platform (2), and the two ends of the length of the generation block (33) are connected to the second working platform (2) via connecting ears, and the generation block (33) is formed with the first generation hole (31) and the second generation hole (32). The droplet generation module (3) also comprises a generation cover plate (34), and the generation cover plate (34) has a covering position for sealing the top surface of the chip oil-liquid hole (101) and the chip gas-liquid hole (102) when generating micro-droplets and realizing the pressure connection between the gas path on the generation cover plate (34) and the chip gas-liquid hole (102).

6. The micro-droplet generation and amplification device according to claim 5, characterized in that: The generating cover plate (34) and the generating pressure block (33) are hingedly connected via a rotating shaft (36); the generating pressure block (33) is provided with an electromagnet (35); when the electromagnet (35) is energized, the generating cover plate (34) is attracted and rotates around the rotating shaft (36) to be in the covering position; and / or, both ends of the length of the generating pressure block (33) are vertically slidably connected to the second working platform (2) via the connecting ears; a guide rod (38) is provided between the top surface of the generating pressure block (33) and the bottom surface of the second working platform (2); a third return spring (39) is sheathed on the guide rod (38); the third return spring (39) is clamped between the bottom surface of the second working platform (2) and the top surface of the generating pressure block (33).

7. The micro-droplet generation and amplification device according to claim 6, characterized in that: The electromagnets (35) are provided with two, the two electromagnets (35) being respectively located at the two ends of the length of the generating pressing block (33), and the generating cover plate (34) is provided with a suction block corresponding to the position of each electromagnet (35) at both ends of the length; and / or, a second return spring (37) is sleeved on the rotating shaft (36) for switching the generating cover plate (34) from the covering position to the opening position when the electromagnet (35) is powered off; and / or, a hand-holding protrusion is provided on an edge of one side of the generating cover plate (34) away from the rotating shaft (36); and / or, a cover plate sealing gasket (310) is provided on an end surface of one side of the generating cover plate (34) facing the generating pressing block (33) and is arranged around each of the generating first holes (31) and the generating second holes (32).

8. The micro-droplet generation and amplification device according to claim 1, characterized in that: The PCR amplification module (4) comprises a semiconductor heating element (41), a first end of the semiconductor heating element (41) being connected to a heat conducting block (43), the temperature control cavity being formed on an end surface of the heat conducting block (43) facing away from the semiconductor heating element (41), and a second end of the semiconductor heating element (41) being connected to a heat dissipation structure.

9. The micro-droplet generation and amplification device according to claim 8, characterized in that: The heat dissipation structure comprises a first heat dissipation block (44) contacting and connected to the second end and a second heat dissipation block (48) contacting and connected to the first heat dissipation block (44); the heat dissipation area of ​​the second heat dissipation block (48) is greater than the heat dissipation area of ​​the first heat dissipation block (44); the heat dissipation structure also comprises a third heat dissipation block (47) and a heat pipe (49); the first end of the heat pipe (49) is located between the first heat dissipation block (44) and the second heat dissipation block (48); and the second end of the heat pipe (49) is connected to the third heat dissipation block (47) for heat exchange.

10. A method for controlling a micro-droplet generation and amplification device according to any one of claims 1 to 9, characterized in that: The steps include: Placing the integrated micro-droplet chip (100) loaded with the sample liquid into the chip placement groove (13); Controlling the second operation platform (2) to switch from a chip picking and placing state to a working state; Controlling the droplet generation module (3) to operate so as to generate microdroplets in the integrated microdroplet chip (100), and storing the microdroplets in a reaction chamber (103) of the integrated microdroplet chip (100); After the micro-droplets are generated, the first working platform (1) and the second working platform (2) are controlled to synchronously flip 180 degrees, and the PCR amplification module (4) is controlled to operate to amplify the micro-droplets in the reaction chamber (103). During the operation of the PCR amplification module (4), the first working platform (1) and the second working platform (2) are controlled to synchronously swing back and forth.

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