Particle pairing device and particle pairing system

Abstract

The utility model discloses a particle pairing device and a pairing system, the pairing device is used for controlling pairing of a first particle and a second particle in a micro-fluidic chip, and the particle pairing device comprises a detection module and a valve control module electrically connected to the detection module, the detection module is used for identifying first particles when a first particle solution flows through a first particle flow channel of the micro-fluidic chip, is also used for identifying second particles when a second particle solution flows through a second particle flow channel of the micro-fluidic chip, and is electrically connected to the valve control module; the valve control module is used for blocking the first particle flow channel when the detection module recognizes the first particles and blocking the second particle flow channel when the detection module recognizes the second particles. Therefore, pairing of a single first particle and a single second particle can be realized, for example, when the first particle and the second particle are respectively cells and microspheres, the accuracy and efficiency of pairing of the cells and the microspheres can be improved, and then the efficiency of single cell sequencing is improved.

Description

Technical Field

[0001] The present application relates to the field of single-cell sequencing technology, and in particular to a particle pairing device and a particle pairing system. Background Art

[0002] Single-cell RNA sequencing (scRNA-seq) reveals the heterogeneity of cells within tissues. Single-cell sequencing is widely used because it offers the advantages of high precision and high resolution over traditional gene sequencing technologies, and can provide information at the level of individual cells. Single-cell sequencing captures genetic information based on a device in which cells and encoded microspheres are co-encapsulated. Currently, a technique uses microvalves to trap cells or microspheres and uses fluids in cell and microsphere channels to push them out, encapsulating them in oil and forming droplets. During this process, multiple microspheres or cells may be encapsulated, and this technique can easily cause contamination of empty droplets, affecting subsequent sequencing data. Furthermore, because the external light beam is prone to refraction and reflection when irradiating the microfluidic chip, the image observed by the operator during the pairing process is cluttered and unclear, making it impossible for the operator to accurately observe and identify cells and microspheres during the pairing operation, further affecting the quality of cell-microsphere pairing. Utility Model Content

[0003] This application aims to address at least one of the technical problems existing in the prior art. To this end, this application proposes a particle pairing device capable of pairing a single first particle with a single second particle to ensure the quality of the particle pairing. For example, when the first particle is a cell and the second particle is a microsphere, respectively, this device can improve the accuracy and efficiency of cell-microsphere pairing, thereby improving the efficiency of single-cell sequencing.

[0004] This application also proposes a particle pairing system and an optical module.

[0005] According to the first embodiment of the present application, a particle pairing device is used to control the pairing of a first particle and a second particle in a microfluidic chip. The microfluidic chip is provided with a pairing area and a first particle flow channel and a second particle flow channel communicating with the pairing area. The particle pairing device includes:

[0006] a detection module, the detection module being configured to identify first particles when the first particle solution flows through the first particle flow channel, and the detection module being configured to identify second particles when the second particle solution flows through the second particle flow channel;

[0007] A valve control module is electrically connected to the detection module, and is used to block the first particle flow channel when the detection module identifies a first particle, and to block the second particle flow channel when the detection module identifies a second particle.

[0008] The particle pairing device according to the embodiment of the present application has at least the following beneficial effects:

[0009] The particle pairing device includes a detection module and a valve control module. By setting the detection module electrically connected to the valve control module, when a first particle solution flows through a first particle flow channel, the detection module can identify the first particle, and at the same time, the valve control module blocks the first particle flow channel to control the number of first particles in the first particle flow channel. When a second particle solution flows through a second particle flow channel, the detection module identifies the second particle, and at the same time, the valve control module blocks the second particle flow channel to control the number of second particles in the second particle flow channel. This can help control the number of first particles and second particles entering the pairing area during the pairing process, so as to achieve pairing of a single first particle and a single second particle in the microfluidic chip, that is, a single droplet only contains a single first particle and a single second particle, thereby ensuring the quality of particle pairing. In this way, for example, when the first particle and the second particle are cells and microspheres, respectively, the accuracy and efficiency of cell-microsphere pairing can be improved, thereby reducing the waste of time and material costs caused by unqualified paired samples, thereby improving the efficiency of single-cell sequencing.

[0010] In other embodiments of the present application, the detection module includes a first optical system, and the particle pairing device also includes an objective lens. The first optical system is used to form a first detection light that passes through the objective lens and illuminates the first particle flow channel, and is used to form a second detection light that passes through the objective lens and illuminates the second particle flow channel. The first particles can be illuminated by the first detection light to emit a first excitation light, and the second particles can be illuminated by the second detection light to emit a second excitation light, wherein the objective lens is configured to at least adjust the spot size of the first detection light and the second detection light.

[0011] In other embodiments of the present application, the light spot is a strip-shaped light spot, and the objective lens is configured to at least adjust the width and / or length of the strip-shaped light spot of the first detection light and the second detection light.

[0012] In other embodiments of the present application, the detection module includes a second optical system, which is configured to receive imaging light from the microfluidic chip and pass through the objective lens to form an image of the microfluidic chip, wherein the objective lens is further configured to magnify the image.

[0013] In other embodiments of the present application, the objective lens is configured such that the first detection light and the second detection light pass through the objective lens in a direction opposite to the direction in which the imaging light passes through the objective lens.

[0014] In other embodiments of the present application, the first optical system includes a first light source and a second light source, the first light source is used to emit the first detection light, the second light source is used to emit the second detection light, and the second optical system includes an image generating component, the image generating component is used to receive the imaging light and form the image;

[0015] The first optical system and the second optical system include a first common optical path, the first common optical path includes a dichroic mirror, and the dichroic mirror is configured to: reflect the first detection light and the second detection light to the objective lens, and transmit the first excitation light, the second excitation light and the imaging light.

[0016] In other embodiments of the present application, the first optical system further includes a detection component, the detection component being configured to receive the first excitation light and the second excitation light, and identify the first particle based on the first excitation light, and identify the second particle based on the second excitation light;

[0017] The first common optical path further includes a first dichroic mirror, which is disposed on the optical path between the dichroic mirror and the detection component, and is configured to: reflect the imaging light emitted from the dichroic mirror to the image generation component, and transmit the first excitation light and the second excitation light emitted from the dichroic mirror to the detection component;

[0018] Alternatively, the first dichroic mirror is configured to transmit the imaging light emitted through the multichromatic mirror to the image generating component, and reflect the first excitation light and the second excitation light emitted through the multichromatic mirror to the detecting component.

[0019] In other embodiments of the present application, the first optical system further includes a detection assembly, the detection assembly including a first detection device and a second detection device, the first detection device being configured to receive the first excitation light and identify the first particles based on the first excitation light, and the second detection device being configured to receive the second excitation light and identify the second particles based on the second excitation light;

[0020] In which, the first optical system also includes a second dichroic mirror, which is arranged on the optical path between the polychroic mirror and the detection component, and the second dichroic mirror is configured to: reflect the first excitation light that passes through the polychroic mirror and is emitted to the first detection device, and transmit the second excitation light that passes through the polychroic mirror and is emitted to the second detection device.

[0021] In other embodiments of the present application, the first common optical path also includes a first reflector, which is arranged on the optical path between the multichromatic mirror and the image generating component, and the first reflector is configured to reflect the first excitation light, the second excitation light and the imaging light that pass through the multichromatic mirror and are emitted.

[0022] In other embodiments of the present application, the objective lens, the multichromatic mirror and the first reflector are arranged in a vertical direction, and the first reflector is configured such that: the first excitation light, the second excitation light and the imaging light that pass through the multichromatic mirror and are emitted are emitted toward the first reflector in a vertical direction, and are emitted in a horizontal direction after being reflected by the first reflector.

[0023] In other embodiments of the present application, the second optical system further includes an image generating component and a quarter-wave plate, wherein the image generating component is used to receive the imaging light and form the image, and the quarter-wave plate is arranged on the optical path between the objective lens and the image generating component.

[0024] In other embodiments of the present application, the first optical system includes a first light source and a second light source, the first light source is used to emit the first detection light, and the second light source is used to emit the second detection light;

[0025] The second optical system further includes a dichroic mirror, which is disposed on an optical path between the objective lens and the image generating assembly, and is configured to: reflect the first detection light and the second detection light to the objective lens, and transmit the first excitation light, the second excitation light, and the imaging light;

[0026] Wherein, the first optical system further includes a polarization beam splitter cube, and the polarization beam splitter cube is arranged on the optical path between the first light source, the second light source and the dichroic mirror.

[0027] In other embodiments of the present application, the first optical system includes a first light source, a second light source and a multichromatic mirror, the first light source is used to emit the first detection light, the second light source is used to emit the second detection light, and the multichromatic mirror is arranged on the optical path between the first light source, the second light source and the objective lens, wherein the multichromatic mirror is configured to: reflect the first detection light and the second detection light to the objective lens, and transmit the first particle to emit the first excitation light after being irradiated by the first detection light, and the second particle to emit the second excitation light after being irradiated by the second detection light.

[0028] In other embodiments of the present application, the first optical system further includes a third dichroic mirror, which is arranged on the optical path between the first light source, the second light source and the multichroic mirror, and the third dichroic mirror is configured to reflect the first detection light and transmit the second detection light.

[0029] In other embodiments of the present application, the first optical system further includes a second reflecting mirror, the second reflecting mirror being disposed on a light path between the second light source and the third dichroic mirror, the second reflecting mirror being configured to reflect the second detection light to the third dichroic mirror;

[0030] The optical axis of the first light source is parallel to the optical axis of the second light source, and the first light source and the second light source are distributed in a horizontal direction.

[0031] In other embodiments of the present application, the wavelengths of the first detection light and the second detection light are different.

[0032] In other embodiments of the present application, the first detection light is perpendicular to the first particle flow channel in the direction from the first particle flow channel to the pairing area, and the second detection light is perpendicular to the second particle flow channel in the direction from the second particle flow channel to the pairing area.

[0033] In other embodiments of the present application, the first detection light and the second detection light enter the objective lens at different incident positions, and / or the first detection light and the second detection light enter the objective lens at different incident angles.

[0034] In other embodiments of the present application, the particle pairing device further includes a supporting platform, the supporting platform is used to mount the microfluidic chip, and the objective lens is disposed on the other side of the supporting platform that is different from the microfluidic chip.

[0035] In other embodiments of the present application, the support platform is provided with a detection window corresponding to the microfluidic chip, and the detection window is connected to two opposite sides of the support platform. Along the height direction of the particle pairing device, the projections of the pairing area, the first particle flow channel and the second particle flow channel are all located within the projection of the detection window, so that the objective lens can observe the microfluidic chip through the detection window.

[0036] In other embodiments of the present application, the particle pairing device further includes a three-axis motion mechanism, the support platform is mounted on the three-axis motion mechanism, and the three-axis motion mechanism is used to drive the support platform to move in three directions perpendicular to each other.

[0037] In other embodiments of the present application, the detection module includes a second optical system, the particle pairing device also includes an objective lens, the second optical system is configured to receive imaging light from the microfluidic chip and passing through the objective lens to form an image of the microfluidic chip, wherein the objective lens is further configured to magnify the image.

[0038] In other embodiments of the present application, the particle pairing device is provided with a liquid reservoir, which stores a first particle solution, a second particle solution, a buffer solution and an oil. The liquid reservoir is used to pass the first particle solution into the first particle flow channel, to pass the second particle solution into the second particle flow channel, and to pass the buffer solution and the oil into the pairing area.

[0039] In other embodiments of the present application, the particle pairing device is provided with a plurality of sample collection tanks and a plurality of sample collection tubes, the plurality of sample collection tanks correspond one-to-one to the plurality of sample collection tubes, and the sample collection tubes are placed in the sample collection tanks, and the sample collection tubes are used to collect droplets generated by the microfluidic chip.

[0040] In other embodiments of the present application, the particle pairing device further includes a control panel, which is electrically connected to the valve control module and is used to control the start and stop of the valve control module.

[0041] In other embodiments of the present application, the valve control module includes at least a first particle flow channel control valve and a second particle flow channel control valve, the first particle flow channel control valve is used to block the first particle flow channel when the detection module identifies the first particle, and the second particle flow channel control valve is used to block the second particle flow channel when the detection module identifies the second particle.

[0042] According to the second embodiment of the present application, the particle pairing system includes a microfluidic chip and the particle pairing device, wherein the microfluidic chip is provided with a pairing area and a first particle flow channel and a second particle flow channel connected to the pairing area, and the microfluidic chip is installed in the particle pairing device.

[0043] According to a third embodiment of the present application, an optical module for identifying a first particle and a second particle includes:

[0044] objective lens;

[0045] a first optical system for forming a first detection light passing through the objective lens, and for forming a second detection light passing through the objective lens, wherein the first particles can be irradiated by the first detection light to emit a first excitation light, and the second particles can be irradiated by the second detection light to emit a second excitation light;

[0046] a second optical system, configured to receive imaging light from the microfluidic chip and passing through the objective lens, to form images of the first particles and / or the second particles;

[0047] The objective lens is configured to adjust the spot sizes of the first detection light and the second detection light, and to amplify the image.

[0048] In other embodiments of the present application, the light spot is a strip-shaped light spot, and the objective lens is configured to at least adjust the width and / or length of the strip-shaped light spot of the first detection light and the second detection light.

[0049] In other embodiments of the present application, the first optical system includes a first light source and a second light source, the first light source is used to emit the first detection light, the second light source is used to emit the second detection light, and the second optical system includes an image generating component, the image generating component is used to receive the imaging light and form the image;

[0050] The first optical system and the second optical system include a first common optical path, the first common optical path includes a dichroic mirror, and the dichroic mirror is configured to: reflect the first detection light and the second detection light to the objective lens, and transmit the first excitation light, the second excitation light and the imaging light.

[0051] In other embodiments of the present application, the first optical system further includes a detection component, the detection component being configured to receive the first excitation light and the second excitation light, and identify the first particle based on the first excitation light, and identify the second particle based on the second excitation light;

[0052] The first common optical path further includes a first dichroic mirror, which is disposed on the optical path between the dichroic mirror and the detection component, and is configured to: reflect the imaging light emitted from the dichroic mirror to the image generation component, and transmit the first excitation light and the second excitation light emitted from the dichroic mirror to the detection component;

[0053] Alternatively, the first dichroic mirror is configured to transmit the imaging light emitted through the multichromatic mirror to the image generating component, and reflect the first excitation light and the second excitation light emitted through the multichromatic mirror to the detecting component.

[0054] In other embodiments of the present application, the first optical system further includes a detection assembly, the detection assembly including a first detection device and a second detection device, the first detection device being configured to receive the first excitation light and identify the first particles based on the first excitation light, and the second detection device being configured to receive the second excitation light and identify the second particles based on the second excitation light;

[0055] In which, the first optical system also includes a second dichroic mirror, which is arranged on the optical path between the polychroic mirror and the detection component, and the second dichroic mirror is configured to: reflect the first excitation light that passes through the polychroic mirror and is emitted to the first detection device, and transmit the second excitation light that passes through the polychroic mirror and is emitted to the second detection device.

[0056] Additional aspects and advantages of the present application will be given in part in the description below, and in part will become obvious from the description below, or will be learned through practice of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] The present application will be further described below with reference to the accompanying drawings and examples, wherein:

[0058] Figure 1 1 is a schematic structural diagram of the particle pairing device disclosed in an embodiment of the present application from one perspective;

[0059] Figure 2 is a schematic structural diagram of the particle pairing device disclosed in an embodiment of the present application from another perspective;

[0060] Figure 3 It is a right side view of the particle pairing device disclosed in the embodiment of the present application;

[0061] Figure 4 Schematic diagram of the optical path of the particle pairing device mainly showing the objective lens disclosed in the embodiment of the present application;

[0062] Figure 5 Schematic diagram of the optical path of the particle pairing device disclosed in the embodiment of the present application mainly showing the detection component;

[0063] Figure 6 Schematic diagram of the optical path of the main display image generation component of the particle pairing device disclosed in the embodiment of the present application;

[0064] Figure 7 This is a light path diagram of the particle pairing device disclosed in the embodiment of the present application, mainly showing the specific structure of the detection component;

[0065] Figure 8 is a light path schematic diagram of the particle pairing device disclosed in an embodiment of the present application, mainly showing a first common light path;

[0066] Figure 9 This is a schematic diagram of the optical path of the particle pairing device disclosed in the embodiment of the present application, mainly showing the first reflector;

[0067] Figure 10 This is a schematic diagram of the optical path of the particle pairing device disclosed in the embodiment of the present application, mainly showing the anti-reflection structure;

[0068] Figure 11 This is a light path diagram of the particle pairing device disclosed in the embodiment of the present application, mainly showing the specific structure of the laser emission mechanism;

[0069] Figure 12 Schematic diagram of the optical path of the particle pairing device disclosed in the embodiment of the present application;

[0070] Figure 13 Schematic diagram of the first laser and the second laser disclosed in the embodiment of the present application being perpendicular to the first particle flow channel and the second particle flow channel respectively;

[0071] Figure 14 A schematic diagram of a microfluidic chip applicable to this application;

[0072] Figure 15 This is a schematic structural diagram of the particle pairing device disclosed in an embodiment of the present application from one perspective, in which the support platform, three-axis motion mechanism, and control panel are hidden;

[0073] Figure 16 for Figure 15 A top view of

[0074] Figure 17 This is a schematic diagram of the connection between the support platform and the three-axis motion mechanism disclosed in the embodiment of this application.

[0075] Reference numerals:

[0076] Particle pairing device 1;

[0077] Microfluidic chip 2, first particle flow channel 21, second particle flow channel 22, oil flow channel 23, buffer flow channel 24, waste liquid flow channel 25, paired flow channel 26, first microvalve 28, second microvalve 29, buffer microvalve 210, waste liquid microvalve 211, oil microvalve 212;

[0078] Detection module 10, laser emitting mechanism 11, first light source 11a, second light source 11b, detection assembly 12, first detection device 12a, second detection device 12b, optical path adjustment mechanism 13, multi-color mirror 1301, first mounting bracket 1302, second mounting bracket 1303, second reflecting mirror 1304, first cylindrical mirror 1305, second cylindrical mirror 1306, third dichroic mirror 1307, third reflecting mirror 1308, fourth reflecting mirror 1309, first reflecting mirror 1310, second dichroic mirror 1311, first filter 1312, second filter 1313, image generation assembly 14, first dichroic mirror 15;

[0079] Valve control module 20;

[0080] Support platform 30, detection window 31, focusing knob 32, liquid reservoir 33, sample collection tank 34, sample collection tube 35, clamp 36;

[0081] Objective lens 40;

[0082] Bright field lamp 50, lamp post 51, bright field light source 52;

[0083] Anti-reflection structure 60, polarization beam splitter cube 61, quarter wave plate 62;

[0084] Three-axis motion mechanism 70, first horizontal drive member 71, second horizontal drive assembly 72, vertical drive assembly 73;

[0085] Control panel 80;

[0086] Base 90, connecting base 91, and column 92. DETAILED DESCRIPTION

[0087] The following describes in detail embodiments of the present application. Examples of the embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present application and are not to be construed as limiting the present application.

[0088] In the description of this application, it should be understood that descriptions involving orientations, such as up, down, front, back, left, right, etc., indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings. They are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation. Therefore, they cannot be understood as limitations on this application.

[0089] In the description of this application, "several" means more than one, "plurality" means more than two, "greater than," "less than," and "exceed" are understood to exclude the number itself, while "above," "below," and "within" are understood to include the number itself. The use of "first" and "second" in the description is solely for the purpose of distinguishing technical features and should not be construed as indicating or implying relative importance, implicitly specifying the number of the indicated technical features, or implicitly specifying the order of the indicated technical features.

[0090] In the description of this application, unless otherwise clearly defined, terms such as setting, installing, and connecting should be understood in a broad sense, and technicians in the relevant technical field can reasonably determine the specific meanings of the above terms in this application based on the specific content of the technical solution.

[0091] In the description of this application, reference to the terms "one embodiment," "some embodiments," "illustrative embodiments," "examples," "specific examples," or "some examples" means that the specific features, structures, materials, or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of this application. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in any appropriate manner in any one or more embodiments or examples.

[0092] The technical solution of the present application will be further described below with reference to the embodiments and drawings.

[0093] Please also refer to Figures 1 to 4 An embodiment of the first aspect of the present application provides a particle pairing device 1 for controlling the pairing of a first particle and a second particle in a microfluidic chip 2. The microfluidic chip 2 is provided with a pairing region and a first particle flow channel and a second particle flow channel communicating with the pairing region. The particle pairing device 1 includes a detection module 10 and a valve control module 20 electrically connected to the detection module 10. The detection module 10 is configured to identify a first particle when a first particle solution flows through the first particle flow channel. The detection module 10 is also configured to identify a second particle when a second particle solution flows through the second particle flow channel. The detection module 10 is electrically connected to the valve control module 20. The valve control module 20 is configured to block the first particle flow channel when the detection module 10 identifies the first particle, and to block the second particle flow channel when the detection module 10 identifies the second particle.

[0094] An embodiment of the present application provides a particle pairing device 1, comprising a detection module 10 and a valve control module 20. The detection module 10 is electrically connected to the valve control module 20, so that when a first particle solution flows through a first particle flow channel, the detection module 10 can identify the first particle, while the valve control module 20 simultaneously blocks the first particle flow channel to control the number of first particles in the first particle flow channel. When a second particle solution flows through a second particle flow channel, the detection module 10 can identify the second particle, while the valve control module 20 simultaneously blocks the second particle flow channel to control the number of second particles in the second particle flow channel. This facilitates controlling the number of first and second particles entering the pairing region during the pairing process, thereby achieving pairing of a single first particle and a single second particle in a microfluidic chip 2, i.e., a single droplet contains only a single first particle and a single second particle, thereby ensuring the quality of the particle pairing. In this way, for example, when the first particle and the second particle are, respectively, a cell and a microsphere, the accuracy and efficiency of cell-microsphere pairing can be improved, thereby reducing the time and material costs wasted due to unqualified paired samples, thereby improving the efficiency of single-cell sequencing.

[0095] Specifically, refer to Figure 14 , shows a front view of a typical microfluidic chip 2, which has multiple flow channels disposed therein, for example, including a first particle flow channel 21, a second particle flow channel 22, an oil flow channel 23, a buffer flow channel 24, a waste liquid flow channel 25, and a pairing flow channel 26. The first particle flow channel 21 is used to allow a first particle solution containing first particles to pass through, the second particle flow channel 22 is used to allow a second particle solution containing second particles to pass through, the buffer flow channel 24 is used to allow a buffer solution to pass through, the oil flow channel 23 is used to allow an oil solution that is incompatible with the first particle solution, the second particle solution, and the buffer solution to pass through, the waste liquid flow channel 25 is used to discharge the first particle solution and the second particle solution, and the pairing flow channel 26 is used to allow a single first particle and a single second particle to reside. The pairing flow channel 26 is connected to the oil flow channel 23, and the first particle flow channel 21, the second particle flow channel 22, and the buffer flow channel 24 are all connected to the pairing flow channel 26.

[0096] The microfluidic chip 2 also includes multiple microvalves, specifically including a pairing microvalve 27 set corresponding to the pairing flow channel 26, a first microvalve 28 set corresponding to the first particle flow channel 21, a second microvalve 29 set corresponding to the second particle flow channel 22, a buffer microvalve 210 set corresponding to the buffer flow channel 24, a waste liquid microvalve 211 set corresponding to the waste liquid flow channel 25, and an oil microvalve 212 corresponding to the oil flow channel 23, wherein the pairing microvalve 27 is used to control the on-off of the pairing flow channel 26, the first microvalve 28 is used to control the on-off of the first particle flow channel 21, the second microvalve 29 is used to control the on-off of the second particle flow channel 22, the buffer microvalve 210 is used to control the on-off of the buffer flow channel 24, the waste liquid microvalve 211 is used to control the on-off of the waste liquid flow channel 25, and the oil microvalve 212 is used to control the on-off of the oil flow channel 23.

[0097] Based on the above structure, the pairing process of the microfluidic chip 2 is as follows:

[0098] The buffer solution microvalve 210 and the paired microvalve 27 are closed, while the first microvalve 28, the second microvalve 29, and the waste liquid microvalve 211 are opened. The first particle solution and the second particle solution are discharged through the paired flow channel 26 and the waste liquid flow channel 25. When a single first particle is identified, the first microvalve 28 is closed, and the single first particle stops in the paired flow channel 26. When a single second particle is identified, the second microvalve 29 is closed, and the single second particle stops in the paired flow channel 26. When both the single first particle and the single second particle stop in the paired flow channel 26, the waste liquid microvalve 211 is closed.

[0099] Based on the above structure, the packaging process of the microfluidic chip 2 is as follows:

[0100] When the first microvalve 28, the second microvalve 29 and the waste liquid microvalve 211 are closed, the buffer microvalve 210 and the matching microvalve 27 are opened, and the buffer transports the single first particle and the single second particle in the matching flow channel 26 into the oil in the oil flow channel 23 to form droplets.

[0101] Based on the embodiment of the first aspect, optionally, the first particles and the second particles can be any one of cells, microspheres or other fine particles, etc., and can be set according to actual needs and are not limited here.

[0102] Based on the embodiment of the first aspect, optionally, the valve control module 20 includes at least a first particle flow channel control valve and a second particle flow channel control valve, the first particle flow channel control valve is used to block the first particle flow channel when the detection module 10 identifies the first particle, and the second particle flow channel control valve is used to block the second particle flow channel when the detection module 10 identifies the second particle.

[0103] In this way, by controlling the first particle flow channel and the second particle flow channel respectively through multiple control valves, the blocking or connection to the outside world of the first particle flow channel and the second particle flow channel can be independently controlled to block the corresponding flow channel when the first particle or the second particle passes through, so that the valve control module 20 can control each flow channel more flexibly, which is conducive to ensuring the pairing of a single first particle and a single second particle.

[0104] Based on the embodiment of the first aspect, optionally, the first particle flow channel control valve and the second particle flow channel control valve can both be any one of pneumatic valves, electric valves, or hydraulic valves, or the first particle flow channel control valve and the second particle flow channel control valve can each be any one of pneumatic valves, electric valves, or hydraulic valves. Exemplarily, the first particle flow channel control valve and the second particle flow channel control valve are both pneumatic valves, so that the valve control module 20 controls the first particle flow channel and the second particle flow channel in a simple, safe, and reliable manner, thereby improving the reliability of the pairing of the first particle and the second particle and ensuring the quality of the particle pairing.

[0105] Based on the embodiment of the first aspect, in some embodiments of the present application, the detection module 10 includes a first optical system, which is used to form a first detection light for irradiating the first particle flow channel, and a second detection light for irradiating the second particle flow channel. The first particle can be irradiated by the first detection light to emit a first excitation light, and the second particle can be irradiated by the second detection light to emit a second excitation light. For example, both the first particle and the second particle have fluorescent dyes, and the first detection light and the second detection light can be lasers, which will generate fluorescence after irradiating the first particle and the second particle. Whether the corresponding particle is detected can be determined by whether the fluorescent signal is captured.

[0106] In this embodiment, refer to Figures 1 to 4 The particle pairing device 1 also includes an objective lens 40. Both the first detection light and the second detection light will pass through the objective lens 40. The objective lens 40 is configured to at least adjust the spot size of the first detection light and the second detection light. Considering that the structural size of the microfluidic chip 2 is small, if the spot area is large, it may cover some inappropriate positions, such as covering another particle flow channel, thereby causing detection errors. That is, the detection module 10 has high requirements for the fineness of the spot when detecting the first particle flow channel and the second particle flow channel. Under the premise of being able to cover the detection position of the corresponding particle flow channel, reducing the area of ​​the spot is conducive to reducing interference. Based on this, the so-called adjustment of the spot size in this embodiment specifically refers to reducing the spot size of the detection module 10 by setting the objective lens 40, so that the detection light of the detection module 10 can accurately irradiate the first particle flow channel and the second particle flow channel, and make the size of the detection light of the detection module 10 adapt to the size of the first particle flow channel and the second particle flow channel, thereby facilitating the detection of the first particle flow channel and the second particle flow channel by the detection module 10.

[0107] In some specific embodiments, the microfluidic chip 2 and the detection module 10 are respectively disposed on the object side (eg, the upper side in the figure) and the image side (eg, the lower side in the figure) of the objective lens 40 .

[0108] When the detection module 10 includes a first optical system, and the first optical system includes an objective lens 40, in some embodiments of the present application, the objective lens 40 can also be used to shape the light spot. Specifically, the cross-section of the light beam emitted by the light source is usually rectangular or circular, and its coverage area is large, and its shape is not suitable for fine flow channels such as the microfluidic chip 2. The objective lens 40 of this embodiment can shape the first detection light and the second detection light so that the light spot irradiated on the microfluidic chip 2 is a strip-shaped light spot (for example, Figure 13 As shown), at this time, the objective lens 40 is configured to at least adjust the corresponding size of the strip light spot, for example, the objective lens 40 is configured to adjust the width of the strip light spot so that the light spot can be concentrated on the detection position. For another example, the objective lens 40 is configured to adjust the length of the strip light spot to adapt to particle flow channels of different widths.

[0109] At the same time, by setting the objective lens 40 to magnify the structure of the microfluidic chip 2, it is also convenient for the operator to observe with the naked eye, so as to realize different detection methods of the second particles and the first particles, and is conducive to the functional inspection of the valve control module 20 and the detection module 10, thereby improving the functional reliability of the particle pairing device 1.

[0110] On the basis of the embodiment of the first aspect, optionally, referring to Figure 1 The particle pairing device 1 further includes a support platform 30, which is disposed on the object side of the objective lens 40. The microfluidic chip 2 is mounted on the support platform 30. The support platform 30 supports the microfluidic chip 2, so that the microfluidic chip 2 remains stable and stationary during operation of the particle pairing device 1, thereby improving the operating reliability of the particle pairing device 1.

[0111] Based on the embodiment of the first aspect, the support platform 30 is optionally provided with a clamp 36 for clamping the microfluidic chip 2. This facilitates stable mounting of the microfluidic chip 2 on the particle pairing device 1, facilitating detection and control of the first and second particle flow channels by the particle pairing device 1. Furthermore, clamping the microfluidic chip 2 by the clamp 36 facilitates removal and installation of the microfluidic chip 2, facilitating replacement, maintenance, and upgrades of the microfluidic chip 2. This allows the particle pairing device 1 to be used with different microfluidic chips 2, thereby improving the practicality of the particle pairing device 1.

[0112] On the basis of the embodiment of the first aspect, optionally, referring to Figure 1The support platform 30 is provided with a detection window 31 corresponding to the microfluidic chip 2. The detection window 31 connects the two opposite sides of the support platform 30, and the microfluidic chip 2 is located on the side of the support platform 30 away from the objective lens 40. Along the height direction of the particle pairing device 1, the projections of the pairing area, the first particle flow channel and the second particle flow channel are all located within the projection of the detection window 31, so that the objective lens 40 can observe the microfluidic chip 2 through the detection window 31, thereby ensuring that the detection module 10 and the valve control module 20 can perform particle identification and channel opening and closing control on the microfluidic chip 2 through the objective lens 40.

[0113] In addition, by setting up the detection window 31, the microfluidic chip 2 and the objective lens 40 can be located on both sides of the support platform 30 respectively, which is conducive to the rational use of space, improves the compactness of the structure, and facilitates the installation and disassembly of the microfluidic chip 2 while ensuring the stability of the structure.

[0114] For example, refer to Figure 1 The support platform 3 includes a flat main structure and is connected to the base 90 via a three-axis motion mechanism 70 described below. A detection window 31 extends through the upper and lower sides of the support platform 3. The microfluidic chip 2 is connected to the support platform 3 via a fixture 36 described below and is located above the detection window 31. The objective lens 40 is connected to the base 90 via a connecting seat, connecting rods, and other structures and is located below the detection window 31.

[0115] The above embodiments describe the function of adjusting the spot size and shaping the spot by the objective lens 40. In other embodiments of the present application, the objective lens 40 can also be applied to the imaging system to assist in amplifying the image. Specifically, the detection module 10 includes a second optical system, which is configured to receive imaging light from the microfluidic chip 2 to form an image of the microfluidic chip 2. In this way, it is convenient for the user to directly check whether the particles have reached the corresponding position in the flow channel. As mentioned above, the structural size of the microfluidic chip 2 is small, and the first particle flow channel, the second particle flow channel, the first particle and the second particle are difficult to see clearly with the naked eye. Therefore, the imaging light of this embodiment will also pass through the objective lens 40 to amplify the image through the objective lens 40, so that the user no longer needs to use other tools to see the first particle flow channel, the second particle flow channel, the first particle and the second particle.

[0116] In this embodiment, the objective lens 40 can be used to adjust the spot area, adjust the spot shape, magnify the image, and other functions, which is beneficial to the detection of particulate matter and can make it convenient for users to check the status of particulate matter. In addition, it can also simplify the structure of the detection module 10 and reduce costs.

[0117] When the detection module 10 further includes a second optical system, in some embodiments of the present application, reference is made to Figure 4The objective lens 40 is configured as follows: the direction in which the first detection light and the second detection light pass through the objective lens 40 is opposite to the direction in which the imaging light passes through the objective lens 40. For example, the first detection light and the second detection light pass through the objective lens 40 from bottom to top, and the imaging light passes through the objective lens 40 from top to bottom. Based on optical principles, for optical devices that can adjust the direction of light after it is incident (such as a convex lens), when light is incident from a certain direction, it will converge, but if it is incident from the opposite direction, it will diverge. By setting the incident directions of the detection light and the imaging light, this embodiment can simultaneously achieve the reduction of the light spot and the enlargement of the image through a single light source element.

[0118] When the detection module 10 includes a first optical system and a second optical system, in some embodiments, referring to Figure 1 and Figure 5 The first optical system includes a laser emitting mechanism 11 and a detection component 12. The laser emitting mechanism 11 is used to emit a first detection light to the first particle flow channel via the objective lens 40 and the detection window 31 in sequence, and can also emit a second detection light to the second particle flow channel. The first detection light is used to make the first particles in the first particle flow channel emit a first excitation light, and the second detection light is used to make the second particles in the second particle flow channel emit a second excitation light. The detection component 12 is used to receive the first excitation light and control the first particle flow channel control valve to block the first particle flow channel. The detection component 12 is also used to receive the second excitation light and control the second particle flow channel control valve to block the second particle flow channel.

[0119] In this way, by setting up the laser emitting mechanism 11, the first particle and the second particle respectively emit the first excitation light and the second excitation light under the action of the laser, so that the detection component 12 controls the operation of the first particle flow channel control valve and the second particle flow channel control valve respectively under the control of the corresponding light beam, thereby realizing independent control of the first particle flow channel and the second particle flow channel, and improving the interception accuracy of the particle pairing device 1 for a single first particle and a single second particle.

[0120] It is understandable that in other embodiments, the detection module 10 may also detect the first particles and the second particles by electrode detection or visual image recognition.

[0121] Reference Figure 6The second optical system includes an image generation component 14, which is used to receive imaging light and form an image. This component facilitates the operator's observation of the operating status of the microfluidic chip 2, improves operational convenience, and facilitates visual detection and manual identification of the first and second particles. This allows the particle pairing device 1 to have different detection methods for the first and second particles, thereby enriching the functionality of the particle pairing device 1. Furthermore, the detection module 10 and the valve control module 20 can be interlocked through visual detection or manual identification, thereby improving the reliability of the particle pairing device 1 in pairing the first and second particles and enhancing the quality of the particle pairing. The image generation component 14 may include a camera and a display screen.

[0122] In other embodiments, the first optical system and the second optical system include a first common optical path. Figure 8 The area surrounded by the long dashed line is roughly marked. The first common optical path specifically refers to the pipeline part shared by the excitation light and the imaging light. Figure 2 、 Figure 3 and Figure 8 The optical system includes a dichroic mirror 1301, which is an optical element capable of transmitting or reflecting light of three or more different wavelengths or wavelength ranges. In this embodiment, the dichroic mirror 1301 is configured to reflect the first detection light and the second detection light toward the objective lens 40, and transmit the first excitation light, the second excitation light, and the imaging light. This enables reflection and transmission of different light types through a single optical element, which improves the reusability and compactness of the structure. It should be noted that the dichroic mirror 1301 can utilize known techniques.

[0123] When the first optical system includes the laser emitting mechanism 11, optionally, referring to Figure 7 The laser emitting mechanism 11 includes a first light source 11a and a second light source 11b. The first light source 11a is used to emit a first detection light to the first particle flow channel through the objective lens 40 and the detection window 31 in sequence, and the first detection light is used to excite the first particles in the first particle flow channel to generate a first excitation light. The second light source 11b is used to emit a second detection light to the second particle flow channel through the objective lens 40 and the detection window 31 in sequence, and the second detection light is used to excite the second particles in the second particle flow channel to generate a second excitation light, and the wavelengths of the first detection light and the second detection light are different.

[0124] In this way, using laser to irradiate the first particles and the second particles can ensure that the light has sufficient energy to ensure that the first particles and the second particles can be activated to emit light beams of sufficient brightness. At the same time, by irradiating the first particle flow channel and the second particle flow channel with two beams of laser light of different wavelengths respectively, the first particles and the second particles can be stimulated to emit light beams of different colors and wavelengths respectively, so that the two photoelectric converters respectively receive the light beams emitted by the first particles and the second particles, which is conducive to the independent detection of the first particles and the second particles.

[0125] Optionally, the wavelengths of the first detection light and the second detection light can be any one of 405nm, 488nm, 532nm, 561nm or 640nm, etc., and can be set specifically according to actual needs. There is no restriction here, but it should be noted that the first detection light and the second detection light need to use lights of different wavelengths respectively. The wavelengths of the first detection light and the second detection light need to be selected in combination with the characteristics of the multi-directional color mirror 1301 and other optical elements of the optical path adjustment mechanism 13. The wavelengths of the first excitation light and the second excitation light are determined according to the wavelengths of the first detection light and the second detection light, and are specifically related to the fluorescence activation of the first particle and the second particle.

[0126] Illustratively, in this embodiment, the wavelength of the first detection light is 488 nm, and the wavelength of the second detection light is 640 nm, thereby meeting the design requirements of different wavelengths of the first detection light and the second detection light, and meeting the conditions that the first detection light excites the first particle to emit the first excitation light and the second detection light excites the second particle to emit the second excitation light.

[0127] It can be understood that, in other embodiments, the first detection light and the second detection light may also be lights of other wavelengths.

[0128] When the first optical system and the second optical system include a first common optical path, in some embodiments of the present application, the first optical system further includes a detection component 12 , and the detection component 12 can be understood with reference to the aforementioned embodiments.

[0129] Reference Figure 1 、 Figure 8The first common optical path also includes a first dichroic mirror 15. The first dichroic mirror 15 is arranged on the optical path between the multi-color mirror 1301 and the detection component 12. The first dichroic mirror 15 is configured to: reflect the imaging light emitted after passing through the multi-color mirror 1301 to the image generation component 14, and transmit the first excitation light and the second excitation light emitted after passing through the multi-color mirror 1301 to the detection component 12. In this way, by providing the first dichroic mirror 15, the first excitation light and the second excitation light can pass through the first dichroic mirror 15 to facilitate subsequent entry into the detection component 12, and the imaging light of other wavelengths can be reasonably utilized to generate an image through the image generation component 14, so as to facilitate observation of the working status of the microfluidic chip 2. It should be noted that the first dichroic mirror 15 can adopt a well-known technology.

[0130] As an alternative to the aforementioned embodiment, the first dichroic mirror 15 is configured to transmit the imaging light that passes through the multichromatic mirror 1301 and is emitted to the image generation component 14, and reflect the first excitation light and the second excitation light that pass through the multichromatic mirror 1301 and are emitted to the detection component 12. That is, the difference between this embodiment and the aforementioned embodiment is that the relationship between transmission and reflection is opposite.

[0131] In the aforementioned embodiment, optionally, the first dichroic mirror 15 can be a long-wave pass dichroic mirror, and the two opposite surfaces of the first dichroic mirror 15 are respectively provided with a splitter film and an anti-reflection film, the image generating component 14 is arranged on the side of the first dichroic mirror 15 provided with the splitter film, and the detection component 12 is arranged on the side of the first dichroic mirror 15 provided with the anti-reflection film, and the cutoff wavelength of the first dichroic mirror 15 is lower than the wavelengths of the first excitation light and the second excitation light, so as to ensure that the first excitation light and the second excitation light can both pass through the first dichroic mirror 15, while the imaging light with a wavelength lower than that of the first dichroic mirror 15 enters the image generating component 14 and generates an image.

[0132] Exemplarily, in this embodiment, the first dichroic mirror 15 is a 490nm long-wave pass dichroic mirror, so that light with a wavelength less than 490nm is reflected and enters the image generating component 14, and the first excitation light and the second excitation light with a wavelength greater than 490nm pass through the first dichroic mirror 15 and are emitted to the detection component 12.

[0133] When the first optical system includes the detection assembly 12, and the first common optical path includes the dichroic mirror 1301, in some embodiments of the present application, reference is made to Figure 1 、 Figure 7The detection component 12 includes a first detection device 12a and a second detection device 12b. The first detection device 12a is electrically connected to the first particle flow channel control valve. The first detection device 12a is used to receive the first excitation light and control the first particle flow channel control valve to block the first particle flow channel. The second detection device 12b is electrically connected to the second particle flow channel control valve. The second detection device 12b is used to receive the second excitation light and control the second particle flow channel control valve to block the second particle flow channel.

[0134] In this way, the first excitation light and the second excitation light are respectively received by the first detection device 12a and the second detection device 12b and the operation of the two control valves are respectively controlled, so that the first excitation light and the second excitation light can be converted into two photoelectric signals through the first detection device 12a and the second detection device 12b respectively, so as to improve the accuracy of the photoelectric signal and avoid the formation of interference signals, thereby helping to improve the accuracy of the control of the first particle flow channel and the second particle flow channel, so as to facilitate the interception control of a single first particle and a single second particle.

[0135] Optionally, the first detection device 12a and the second detection device 12b can be any one of photomultiplier tubes (PMT), avalanche photodiodes (APD) or photodiodes, and can be specifically configured according to actual needs and are not limited here.

[0136] In this embodiment, refer to Figure 1 、 Figure 7 、 Figure 8 The first optical system also includes a second dichroic mirror 1311, which is arranged on the optical path between the multichromatic mirror 1301 and the detection component 12. The second dichroic mirror 1311 is configured to: reflect the first excitation light that passes through the multichromatic mirror 1301 and is emitted to the first detection device 12a, and transmit the second excitation light that passes through the multichromatic mirror 1301 and is emitted to the second detection device 12b.

[0137] Specifically, the second dichroic mirror 1311 is used to change the optical paths of the first excitation light and the second excitation light, so that the first excitation light enters the first detection device 12a through the objective lens 40, the multichromatic mirror 1301 and the second dichroic mirror 1311 in sequence, and the second excitation light enters the second detection device 12b through the objective lens 40, the multichromatic mirror 1301 and the second dichroic mirror 1311 in sequence.

[0138] In this way, the first excitation light and the second excitation light have different transmission directions through the second dichroic mirror 1311, so that the two light beams can enter the first detection device 12a and the second detection device 12b respectively, thereby controlling the operation of the first particle flow channel control valve and the second particle flow channel control valve respectively, reducing the signal interference between the two light beams, avoiding signal crossing, and thereby improving the control accuracy of the first particle flow channel and the second particle flow channel.

[0139] Optionally, the second dichroic mirror 1311 can be a long-wave pass dichroic mirror, and the two opposite surfaces of the second dichroic mirror 1311 are respectively provided with a splitter film and an anti-reflection film. The first detection device 12a is arranged on the side of the second dichroic mirror 1311 provided with the splitter film, and the second detection device 12b is arranged on the side of the second dichroic mirror 1311 provided with the anti-reflection film, and the cutoff wavelength of the second dichroic mirror 1311 is between the wavelengths of the first excitation light and the second excitation light, so that the light beam with a wavelength higher than the cutoff wavelength passes through the second dichroic mirror 1311, and the light beam with a wavelength lower than the cutoff wavelength is reflected by the second dichroic mirror 1311, so that the first excitation light and the second excitation light are transmitted in two different directions, so as to satisfy the first excitation light entering the first detection device 12a and the second excitation light entering the second detection device 12b.

[0140] Illustratively, in this embodiment, the second dichroic mirror 1311 is a 550nm long-wave pass dichroic mirror, so that the first excitation light with a wavelength less than 550nm is reflected and enters the first detection device 12a, and the second excitation light with a wavelength greater than 550nm passes through the second dichroic mirror 1311 and enters the second detection device 12b.

[0141] When the second optical system includes the image generating component 14 and the first common optical path includes the dichroic mirror 1301, in some embodiments of the present application, reference is made to Figure 3 、 Figure 9 The first shared optical path also includes a first reflector 1310. The first reflector 1310 is disposed on the optical path between the dichroic mirror 1301 and the image generation assembly 14. In other words, the dichroic mirror 1301 is disposed between the objective lens 40 and the first reflector 1310 along the optical axis of the objective lens 40. The first reflector 1310 is configured to reflect the first excitation light, the second excitation light, and the imaging light emitted after passing through the dichroic mirror 1301. By providing the first reflector 1310 to adjust the transmission direction of the first excitation light and the second excitation light, the size of the particle pairing device 1 along the optical axis of the objective lens 40 can be reduced.

[0142] When the first common optical path further includes a first reflector 1310, in some embodiments of the present application, Figure 3 、 Figure 9The objective lens 40, the pleochroic mirror 1301, and the first reflector 1310 are arranged in a vertical direction. For example, the objective lens 40, the pleochroic mirror 1301, and the first reflector 1310 are arranged in a top-to-bottom direction. The first reflector 1310 is configured such that the first excitation light, the second excitation light, and the imaging light emitted after passing through the pleochroic mirror 1301 are emitted vertically toward the first reflector 1310, and are then emitted horizontally after being reflected by the first reflector 1310. This facilitates placement of the microfluidic chip 2 at the top, facilitating user operation, while fully utilizing the vertical and horizontal space, thereby helping to control the volume of the particle pairing device 1.

[0143] It should be noted that when the first common optical path includes a multi-directional mirror 1301, a first reflector 1310, a first dichroic mirror 15 and a second dichroic mirror 1311, it adopts the following layout: the objective lens 40 is located in the upper layer, the multi-directional mirror 1301 is located in the middle layer, the first reflector 1310, the first dichroic mirror 15 and the second dichroic mirror 1311 are located in the lower layer, and the multi-directional mirror 1301, the first reflector 1310, the first dichroic mirror 15 and the second dichroic mirror 1311 are arranged in sequence along the emission direction of the excitation light.

[0144] Considering that the substrate of the microfluidic chip 2 is a transparent plate, the laser is likely to form a strong reflection when it is irradiated to the microfluidic chip 2 through the detection window 31, resulting in uneven light intensity and unclear images collected by the image generation component 14. It is difficult for the operator to clearly observe the working status of the microfluidic chip 2, and therefore it is impossible to accurately identify the number of cells and microspheres passing through the flow channel, thereby affecting the quality of cell-microsphere pairing. Based on this, when the detection module 10 further includes a second optical system, in some embodiments of the present application, reference is made to Figure 3 、 Figure 10 The second optical system further includes an image generating assembly 14 and a quarter-wave plate 62. The image generating assembly 14 is configured to receive imaging light and form an image, as can be understood with reference to the aforementioned embodiments. The quarter-wave plate 62 is disposed in the optical path between the objective lens 40 and the image generating assembly 14. The quarter-wave plate 62 attenuates light reflected from the transparent plate, thereby reducing stray light collected by the image generating assembly 14 and improving image purity and clarity. This, in turn, improves the accuracy of cell and microsphere identification, thereby enhancing the quality of cell-microsphere pairing.

[0145] Specific to Figure 10 In the illustrated embodiment, when the first common optical path includes the dichroic mirror 1301 , the quarter-wave plate 62 is disposed on the optical path between the objective lens 40 and the dichroic mirror 1301 .

[0146] When the second optical system further includes a quarter wave plate 62, in some embodiments of the present application, referring to Figure 2、 Figure 10 The first optical system includes a first light source 11a and a second light source 11b. The first light source 11a is used to emit a first detection light, and the second light source 11b is used to emit a second detection light. The second optical system also includes a dichroic mirror 1301. The dichroic mirror 1301 is disposed on the optical path between the objective lens 40 and the image generation component 14. The dichroic mirror 1301 is configured to reflect the first detection light and the second detection light toward the objective lens 40 and transmit the first excitation light, the second excitation light, and the imaging light. The first light source 11a, the second light source 11b, and the dichroic mirror 1301 can all be understood with reference to the aforementioned embodiments.

[0147] In this embodiment, the first optical system further includes a polarization beam splitter cube 61, which is disposed in the optical path between the first light source 11a, the second light source 11b and the dichroic mirror 1301. The polarization beam splitter cube 61 can cooperate with the quarter-wave plate 62 to further attenuate the reflected light of the transparent plate.

[0148] The aforementioned embodiments propose that the objective lens can achieve functions such as adjusting the spot area, adjusting the spot shape, and magnifying the image, and propose solutions for a first optical system and a second optical system based on this. Other embodiments of the present application also propose solutions for the first optical system when the detection module does not include a second optical system capable of imaging. Specifically, the first optical system includes a first light source 11a, a second light source 11b, and a dichroic mirror 1301. The first light source 11a is used to emit a first detection light, and the second light source 11b is used to emit a second detection light. The dichroic mirror 1301 is disposed on the optical path between the first light source 11a, the second light source 11b, and the objective lens 40. The dichroic mirror 1301 is configured to reflect the first detection light and the second detection light to the objective lens 40, and transmit the first particle to emit a first excitation light after being irradiated by the first detection light, and the second particle to emit a second excitation light after being irradiated by the second detection light. The first light source 11a, the second light source 11b, and the dichroic mirror 1301 of this embodiment can all be understood with reference to the aforementioned embodiments.

[0149] Based on the above embodiments, in some embodiments of the present application, refer to Figure 2 、 Figure 11 The first optical system also includes a third dichroic mirror 1307, which is positioned on the optical path between the first and second light sources 11a, 11b, and the dichroic mirror 1301. This mirror is configured to reflect the first detection light and transmit the second detection light. This third dichroic mirror 1307 adjusts the laser direction so that the two detection light beams emitted by the first and second light sources 11a, 11b are emitted in the same direction, allowing both the first and second detection light beams to enter the objective lens 40.

[0150] When the first optical system also includes a third dichroic mirror 1307, the first optical system also includes a second reflector 1304, and the second reflector 1304 is arranged on the optical path between the second light source 11b and the third dichroic mirror 1307. The second reflector 1304 is configured to reflect the second detection light to the third dichroic mirror 1307. By setting the second reflector 1304, the first detection light passes through an optical element more than the second detection light to change the direction of the first detection light, which is conducive to making the first detection light and the second detection light enter the objective lens 40 in different directions, thereby facilitating a more flexible layout of the first light source 11a and the second light source 11b.

[0151] Based on the above structure, the light source of this embodiment can adopt the following layout: the optical axis of the first light source 11a is parallel to the optical axis of the second light source 11b, and the first light source 11a and the second light source 11b are distributed in the horizontal direction. In this way, the first light source 11a and the second light source 11b are located on one side of the third dichroic mirror 1307 and the second reflector 1304. Specifically, the optical axes of the first light source 11a and the second light source 11b are perpendicular to the optical axes of the third dichroic mirror 1307 and the second reflector 1304, thereby making the layout of the optical device more compact and helping to reduce the space occupied by the first optical system.

[0152] When the particle pairing device further includes an objective lens 40, optionally, the first detection light (see Figure 13 S1 in the figure) is perpendicular to the first particle flow channel (the first detection light is perpendicular to the page at this time), and the second detection light (see Figure 13 S2 in the figure is perpendicular to the second particle flow channel (the second detection light is perpendicular to the page at this time). This is beneficial to ensure that the first particle can be illuminated by the first detection light when passing through the first particle flow channel, and ensure that the second particle can be illuminated by the second detection light when passing through the second particle channel, thereby improving the detection accuracy of the detection module 10 for the first particle and the second particle, and further improving the reliability of particle pairing.

[0153] When the particle pairing device further includes an objective lens 40, optionally, the first detection light and the second detection light enter the objective lens 40 at different incident positions, and the first detection light does not intersect with the second particle flow channel, and the second detection light does not intersect with the first particle flow channel, thereby ensuring that the first particle is only activated by the first detection light and emits the first excitation light, and the second particle is only activated by the second detection light and emits the second excitation light, thereby facilitating reduction of signal interference and avoiding the situation where the first particle is simultaneously affected by the first detection light and the second detection light, thereby further improving the accuracy of the particle pairing device 1 in detecting the first and second particles. More specifically, the first detection light and the second detection light both enter the objective lens 40 in a direction parallel to the optical axis of the objective lens 40.

[0154] When the particle pairing device further includes an objective lens 40, optionally, the first detection light and the second detection light enter the objective lens 40 at different incident angles, and the first detection light and the second particle flow channel do not intersect, and the second detection light and the first particle flow channel do not intersect, to ensure that the first particle is only activated by the first detection light and emits the first excitation light, and the second particle is only activated by the second detection light and emits the second excitation light, so as to help reduce signal interference and avoid the situation where the first particle is simultaneously affected by the first detection light and the second detection light and generates an interference peak, thereby further improving the accuracy of the particle pairing device 1 in detecting the first and second particles. It should be noted that in this case, the positions at which the first detection light and the second detection light enter the objective lens 40 can be the same or different.

[0155] When the particle pairing device also includes an objective lens 40, in some embodiments, the detection module 10 also includes an optical path adjustment mechanism 13, which is arranged between the objective lens 40 and the laser emitting mechanism 11 and the detection component 12. The optical path adjustment mechanism 13 is used to adjust the direction and shape of the first detection light and the second detection light, and to adjust the direction of the first excitation light and the second excitation light, so as to improve the structural compactness while ensuring that the first detection light and the second detection light are irradiated to the first particle flow channel and the second particle flow channel, and the first excitation light and the second excitation light are received by the first detection device 12a and the second detection device 12b, so as to facilitate reducing the volume of the particle pairing device 1.

[0156] Optionally, the optical path adjustment mechanism 13 includes the aforementioned multi-directional chromatic mirror 1301, which is arranged on the side of the objective lens 40 away from the detection window 31. The multi-directional chromatic mirror 1301 is used to reflect the laser emitted by the laser emitting mechanism 11 to the objective lens 40 and to pass the first excitation light and the second excitation light, thereby ensuring that the first excitation light, the second excitation light, the first detection light and the second detection light are all within the focusing or imaging range of the objective lens 40. At the same time, it can also help to improve the reusability of the structure and further improve the compactness of the structure.

[0157] Optionally, refer to Figure 2 The optical path adjustment mechanism 13 also includes a first mounting bracket 1302 and a second mounting bracket 1303, which are spaced apart. The first mounting bracket 1302 is used to mount the first light source 11a and adjust the emission direction of the first detection light. The second mounting bracket 1303 is used to mount the second light source 11b and adjust the emission direction of the second detection light. This facilitates adjusting the directions in which the first and second detection lights enter the objective lens, ensuring that the first and second detection lights enter the objective lens 40 at different angles of incidence.

[0158] Furthermore, the first mounting bracket 1302 and the second mounting bracket 1303 can structurally adjust the laser emission directions of the first light source 11a and the second light source 11b to change the emission directions of the first detection light and the second detection light from the laser generator, or can change the emission directions of the first detection light and the second detection light through the first mounting bracket 1302 and the second mounting bracket 1303 by setting optical elements internally. The specific settings can be made according to actual needs and are not limited here.

[0159] Optionally, the optical path adjustment mechanism 13 further includes the aforementioned second reflector 1304 . The second reflector 1304 is disposed in the emitting direction of the first detection light of the first light source 11 a . The second reflector 1304 is used to adjust the optical path direction of the first detection light.

[0160] Thus, by providing the second reflector 1304 , the first detection light passes through an additional optical element compared to the second detection light to change its direction, thereby facilitating the first detection light and the second detection light to enter the objective lens 40 in different directions.

[0161] Optionally, refer to Figure 2 、 Figure 11 The optical path adjustment mechanism 13 further includes a first cylindrical mirror 1305 and a second cylindrical mirror 1306. The first cylindrical mirror 1305 is disposed between the second reflector 1304 and the dichroic mirror 1301, and the second cylindrical mirror 1306 is disposed between the second mounting bracket 1303 and the dichroic mirror 1301. The first cylindrical mirror 1305, the second cylindrical mirror 1306, and the objective lens 40 convert the first and second detection lights into stripe lasers to adjust the image heights of the first and second detection lights, thereby adjusting the illumination areas of the first and second detection lights, thereby facilitating detection and identification of the first and second particles in a specified area.

[0162] Optionally, the optical path adjustment mechanism 13 also includes the aforementioned third dichroic mirror 1307, which is arranged between the first cylindrical mirror 1305, the second cylindrical mirror 1306 and the multi-directional mirror 1301. The direction of the laser passing through the cylindrical mirror is adjusted by the third dichroic mirror 1307 so that both the first detection light and the second detection light can enter the objective lens 40.

[0163] Optionally, refer to Figure 2The optical path adjustment mechanism 13 further includes a third reflector 1308 and a fourth reflector 1309. The third reflector 1308 and the fourth reflector 1309 are sequentially arranged between the third dichroic mirror 1307 and the multi-directional mirror 1301. The optical path directions of the first detection light and the second detection light are folded back again by the third reflector 1308 and the fourth reflector 1309, so as to facilitate adjusting the direction of the laser entering the objective lens 40 and reduce the size of the particle pairing device 1. Specifically, the first light source 11a, the second light source 11b, the third dichroic mirror 1307 and the third reflector 1308 are all located at the bottom layer, and the fourth reflector 1309 and the multi-directional mirror 1301 are located in the middle layer, that is, located above the first light source 11a, the second light source 11b, the third dichroic mirror 1307 and the third reflector 1308. Specifically, the fourth reflector 1309 is located directly above the third reflector 1308, and the objective lens 40 is located in the upper layer, specifically at the multi-directional mirror 1301. Directly above the dichroic mirror 1301, the detection light passes through the third dichroic mirror 1307 and is incident on the third reflector 1308. After being reflected by the third reflector 1308, it is incident upward on the fourth reflector 1309. After being reflected by the fourth reflector 1309, it is incident on the multi-color mirror 1301 on the same layer. After being reflected by the multi-color mirror 1301, it is incident upward on the objective lens 40. In this way, the horizontal and vertical spaces can be fully utilized to arrange various optical devices, thereby improving space utilization.

[0164] Optionally, the optical path adjustment mechanism 13 also includes a polychromatic narrow-band filter, which is arranged between the polychromatic mirror 1301 and the fourth reflector 1309. The polychromatic narrow-band filter filters light beams of some wavelengths to ensure the purity of the light beam entering the polychromatic mirror 1301, thereby helping to improve the purity of the light beam irradiated to the first particle flow channel and the second particle flow channel, so as to facilitate the detection of the first particles and the second particles.

[0165] Optionally, refer to Figure 4 、 Figure 12 A first filter 1312 is provided at the light inlet of the first detection device 12a, and a second filter 1313 is provided at the light inlet of the second detection device 12b, so as to limit the passage of light beams of a specific wavelength and filter out other stray light, thereby helping to improve the purity of the photoelectric signal.

[0166] Exemplarily, the first filter 1312 is a 525nm narrowband filter, and the second filter 1313 is a 655nm longwave pass filter, so that the light beam with a wavelength near 525nm enters the first detection device 12a, and the light beam with a wavelength longer than 655nm enters the second detection device 12b.

[0167] It is understandable that in other embodiments, the second dichroic mirror 1311 and the first dichroic mirror 15 may also be short-wave pass dichroic mirrors to allow light beams with wavelengths lower than the cutoff wavelength to pass through and light beams with wavelengths higher than the cutoff wavelength to be reflected.

[0168] In some embodiments, referring to Figure 1 The particle pairing device 1 further includes a brightfield lamp 50 for illuminating the detection window 31. The brightfield lamp 50 is disposed on a side of the detection window 31 away from the objective lens 40, with the brightfield lamp 50 and the objective lens 40 located on opposite sides of the detection window 31. This improves structural rationality and facilitates reducing the size of the particle pairing device 1. Light emitted by the brightfield lamp 50 sequentially passes through the detection window 31, the objective lens 40, the multi-color mirror 1301, the first reflector 1310, and the first dichroic mirror 15, entering the image generation component 14, thereby illuminating the surface of the microfluidic chip 2. This facilitates the introduction of brightfield light into the image generation component 14, thereby increasing the brightness of the image generated by the image generation component 14 and improving the imaging quality, thereby facilitating the operator's observation of the microfluidic chip 2.

[0169] Optionally, the wavelength of the light emitted by the bright field lamp 50 is different from the wavelengths of the first excitation light and the second excitation light, which is conducive to distinguishing the light of the bright field lamp 50 from the first excitation light and the second excitation light and driving light of different wavelengths into different optical elements to act, thereby avoiding the situation where the light of the bright field lamp 50 mistakenly enters the first detection device 12a or the second detection device 12b, resulting in incorrect detection of the first particle and the second particle.

[0170] Furthermore, as can be seen from the foregoing, the second dichroic mirror 1311 is a 550nm long-wave pass dichroic mirror. Based on this, the wavelength of the light beam emitted by the bright field lamp 50 can be 445nm, 448nm or 450nm, etc., so as to meet the condition of being reflected by the second dichroic mirror 1311 to ensure that it can enter the image generation component 14.

[0171] Optionally, a narrowband filter is provided at the light outlet of the brightfield lamp 50, which can help filter the light emitted by the brightfield lamp 50 so that the wavelength of the light emitted through the brightfield lamp 50 and the narrowband filter is in the target band, so as to help reduce the impact of the light of the brightfield lamp 50 on the particle detection results.

[0172] For ease of reading and understanding, the following Figure 12 A simple example is given to illustrate the motion path of the light beam of the particle pairing device 1:

[0173] The wavelength of the first detection light is 488 nm, and the wavelength of the second detection light is 640 nm. After the first detection light is emitted by the first light source 11 a, it passes through the first mounting bracket 1302, the second reflector 1304, the first cylindrical mirror 1305, the third dichroic mirror 1307, the third reflector 1308, the fourth reflector 1309, the polarization beam splitting cube 61, the polychromatic narrow-band filter, the polychromatic mirror 1301, the quarter-wave plate 62, the objective lens 40 and the detection window 31 to enter the microfluidic chip 2. After the second detection light is emitted by the second light source 11 b, it passes through the second mounting bracket 1303, the second cylindrical mirror 1306, the third dichroic mirror 1307, the third reflector 1308, the fourth reflector 1309, the polarization beam splitting cube 61, the polychromatic narrow-band filter, the polychromatic mirror 1301, the quarter-wave plate 62, the objective lens 40 and the detection window 31 to enter In the microfluidic chip 2, the first detection light excites the first particle to emit the first excitation light, and the second detection light excites the second particle to emit the second excitation light. The bright field light emitted by the bright field lamp 50 and the first excitation light and the second excitation light pass through the detection window 31, the objective lens 40, the quarter-wave plate 62, the multi-directional color mirror 1301 and the first reflecting mirror 1310 in sequence, so that the bright field light, the first excitation light and the second excitation light are reflected to the first dichroic mirror 15. At this time, the bright field light is reflected by the first dichroic mirror 15 to enter the image generating component 14, the first excitation light and the second excitation light pass through the first dichroic mirror 15, and then the first excitation light is reflected by the second dichroic mirror 1311 to the first filter 1312, and passes through the first filter 1312 to enter the first detection device 12a, and the second excitation light passes through the second dichroic mirror 1311 and the second filter 1313 in sequence to enter the second detection device 12b.

[0174] For ease of understanding, refer to Figures 14 to 17 , and combined with Figures 1 to 3 The particle pairing device 1 according to the present application will be described.

[0175] Reference Figures 15 to 17 Combined with Figure 1In some embodiments, the particle pairing device 1 further includes a three-axis motion mechanism 70, on which the support platform 30 is mounted. The three-axis motion mechanism 70 is configured to drive the support platform 30 to move relative to the objective lens 40 in three mutually perpendicular directions. This facilitates adjusting the observation position of the objective lens 40 relative to the microfluidic chip 2 and adjusting the focal length of the objective lens 40, thereby improving the imaging quality of the image generation component 14. The three-axis motion mechanism 70 can specifically drive the support platform 30 to move along the first horizontal direction (for example, the front-to-back direction), the second horizontal direction (for example, the left-to-right direction) and the vertical direction. Exemplarily, the three-axis motion mechanism 70 includes a first horizontal drive component 71, a second horizontal drive component 72 and a vertical drive component 73. The first horizontal drive component 71 is fixedly connected to the base 90, the second horizontal drive component 72 is connected to the drive end of the first horizontal drive component 71, the vertical drive component 73 is connected to the drive end of the second horizontal drive component 72, and the support platform 30 is connected to the drive end of the vertical drive component 73. The first horizontal drive component 71, the second horizontal drive component 72 and the vertical drive component 73 can be drive components including a motor, a screw and a screw nut.

[0176] Optionally, refer to Figure 1 and Figure 17 The support platform 30 further includes a focus knob 32 for adjusting the focal length of the objective lens 40, thereby facilitating improvement of the imaging quality of the image generating assembly 14. Exemplarily, the focus knob 32 extends from the upper side of the support platform 30 for easy operation by the user.

[0177] Optionally, refer to Figure 1 The support platform 30 is provided with a liquid reservoir 33, which stores a first particle solution, a second particle solution, a buffer solution and an oil. The liquid reservoir 33 is used to introduce the first particle solution into the first particle flow channel, the second particle solution into the second particle flow channel, and the buffer solution and the oil into the pairing area.

[0178] In this way, by setting up a liquid storage tank 33 to store the solution required in the process of pairing the first particles and the second particles, it can be helpful to shorten the liquid supply path to the microfluidic chip 2, so as to improve the liquid supply efficiency of the microfluidic chip 2. At the same time, it can also make the structure of the particle pairing device 1 more complete, and the particle pairing device 1 can be used independently without the solution raw material pool, which can help improve the portability of the particle pairing device 1.

[0179] Optionally, refer to Figure 2 The support platform 30 is provided with a plurality of sample collection slots 34 and a plurality of sample collection tubes 35. The plurality of sample collection slots 34 correspond one to one with the plurality of sample collection tubes 35, and the sample collection tubes 35 are placed in the sample collection slots 34. The sample collection tubes 35 are used to collect droplets generated by the microfluidic chip 2.

[0180] In this way, the storage of droplets generated after the pairing of the first particle and the second particle can be facilitated, so as to shorten the time for operators to take and place droplets, thereby helping to improve the efficiency of single-cell sequencing.

[0181] It should be noted that the above embodiment uses the support platform 30 as the installation base to install components such as the microfluidic chip 2, focusing knob 32, liquid reservoir 33, collection tank 34 and multiple sample collection tubes 35, which can achieve integrated installation and facilitate user operation.

[0182] In some embodiments, referring to Figure 1 The particle pairing device 1 further includes a control panel 80, which is electrically connected to the first and second particle flow channel control valves of the valve control module 20. The control panel 80 is used to control the start and stop of the first and second particle flow channel control valves. This allows for manual control of the first and second particle flow channel control valves, enabling autonomous adjustment of the valve control module 20 in special circumstances (such as when the detection module 10 is unavailable or the operator has special control requirements for the control valves), thereby improving the flexibility of the particle pairing device 1. Furthermore, by controlling the start and stop of the valve control module 20 through the control panel 80, it can also be used to detect whether the valve control module 20 is functioning properly, thereby improving the reliability of single cell and single microsphere pairing.

[0183] Optionally, the valve control module 20 also includes a control circuit board, and the first light source 11a, the second light source 11b, the first particle flow channel control valve and the second particle flow channel control valve are respectively electrically connected to the control circuit board. The control panel 80 includes a display screen and control buttons, and the display screen and the control buttons are respectively electrically connected to the control circuit board. The control circuit board is arranged on the side of the display surface of the control panel 80 away from the display screen. The display screen is used to display the working data of the valve control module 20 and the detection results of the detection module 10 in real time, such as the working status or air pressure value of the first particle flow channel control valve, etc. There are multiple control buttons, and the multiple control buttons are respectively electrically connected to the first particle flow channel control valve and the second particle flow channel control valve through the control circuit board. The control buttons are used to control the switch and pressure of the first particle flow channel control valve and the second particle flow channel control valve, etc., which is conducive to improving the flexibility of the valve control module 20, so that the particle pairing device 1 can freely adjust parameters according to user needs to improve the practicality of the particle pairing device 1. In addition, multiple control buttons can also be used to adjust the gain of the first detection device 12a and the second detection device 12b, as well as to realize functions such as manually triggering the detection module 10 and the droplet wrapping program. The specific settings can be made according to actual needs and are not limited here.

[0184] Optionally, the control panel 80 further includes an automatic focusing button, which is used to adjust the focal length of the objective lens 40 through electrical control to achieve an automatic focusing function, thereby facilitating improvement in imaging clarity of the image generating component 14 .

[0185] In some embodiments, referring to Figure 1 The particle pairing device 1 also includes a shell and a base 90 arranged at the bottom of the shell. The three-axis motion mechanism 70, the detection module 10 and the valve control module 20 are all arranged on the base 90 and located inside the shell. The control panel 80 is installed in the shell and is at least partially located outside the shell. Multiple structures of the detection module 10 are arranged around the three-axis motion mechanism 70, so that space can be reasonably utilized, which is beneficial for ensuring the volume of the particle pairing device 1 while allowing the detection module 10 to have a longer optical path to facilitate projection imaging of the laser and light beam.

[0186] In some embodiments, referring to Figure 15 The objective lens 40 is fixedly connected to the base 90 via a connecting structure, which includes a connecting seat 91 and a column 92. The bottom end of the column 92 is fixedly connected to the base 90, and the connecting seat 91 is connected to the top of the column 92. Exemplarily, the connecting seat 91 is located lateral to the top of the column 92 and is connected to the top of the column 92 via a horizontally arranged bracket. The objective lens 40 is mounted on the upper side of the connecting seat 91, and the connecting seat 91 is provided with a through hole for light to pass through, which corresponds to the objective lens 40. In other embodiments, the mounting seat 91 is also used to connect to the dichroic mirror 1301. Exemplarily, the dichroic mirror 1301 is connected to the lower side of the mounting seat 91. In other embodiments, the column 92 is also used to mount the brightfield lamp 50. Exemplarily, the brightfield lamp 50 includes a lamp post 51 and a brightfield light source 52. The lamp post 51 is connected to the top of the column 92, and the brightfield light source 52 is connected to the top of the lamp post 51 and is located to the side of the lamp post 52 so that the axis of the brightfield light source 51 remains coaxial with the objective lens 40. In some embodiments, the connector 91 is also used to connect to a quarter-wave plate 62. Exemplarily, the quarter-wave plate 62 is connected to the upper side of the connector 91, and the objective lens 40 is connected to the upper side of the quarter-wave plate 62. That is, the objective lens 40 is connected to the connector 91 via the quarter-wave plate 62.

[0187] In some embodiments, referring to Figure 15 、 Figure 16The laser emitting mechanism 11, the second reflecting mirror 1304, the first cylindrical mirror 1305, the second cylindrical mirror 1306, the third dichroic mirror 1307, the third reflecting mirror 1308, the fourth reflecting mirror 1309, and the multi-directional color mirror 1301 are arranged around the three-axis motion mechanism 70. The first light source 11a, the second light source 11b, the second reflecting mirror 1304, the first cylindrical mirror 1305, the second cylindrical mirror 1306, the third dichroic mirror 1307, the third reflecting mirror 1308, the first detection device 12a, the second detection device 12b, the first reflecting mirror 1310, the second dichroic mirror 1311 and the first dichroic mirror 15 are located at the same height, and all or part of them are connected to the base 90 through connecting columns arranged in the vertical direction, so as to serve as the bottom component of the optical path structure of the particle pairing device 1. The fourth reflector 1309, the polarization beam splitting cube 61, and the multi-color mirror 1301 are located at the same height, and the fourth reflector 1309 is connected above the third reflector 1308. The multi-color mirror 1301 is connected above the first reflector 1310 to serve as a middle-layer component of the optical path structure of the particle pairing device 1. The objective lens 40 is connected to the upper side of the multi-color mirror 1301 to serve as an upper-layer component of the optical path structure of the particle pairing device 1.

[0188] Based on the first embodiment, in some embodiments, the detection module 10 includes a second optical system, and the particle pairing device 1 also includes an objective lens 40. The second optical system is configured to receive imaging light from the microfluidic chip 2 and pass through the objective lens 40 to form an image of the microfluidic chip 2, wherein the objective lens 40 is also configured to magnify the image. The second optical system and the objective lens 40 in this embodiment can be understood with reference to the aforementioned embodiments. This embodiment mainly protects the solution of using the objective lens 40 to magnify the image function.

[0189] In the second aspect, the present application also provides a particle pairing system (not shown), comprising a microfluidic chip 2 and a particle pairing device 1 as described in the first aspect above, the microfluidic chip 2 being provided with a pairing area and a first particle flow channel and a second particle flow channel connecting the pairing area, and the microfluidic chip 2 being installed on the particle pairing device 1.

[0190] It can be understood that, since the particle pairing system includes the particle pairing device 1 described in the first aspect, the particle pairing system has the beneficial effects of the particle pairing device 1 described in the first aspect, which will not be described in detail here.

[0191] For ease of reading and understanding, the following simple example illustrates the workflow of the particle pairing system, assuming the first particle is a cell and the second particle is a microsphere:

[0192] Before the particle pairing system starts working, the microfluidic chip 2 is first mounted on the support platform 30 by means of the clamp 36, the air in the flow channel of the microfluidic chip 2 (including the first particle flow channel, the second particle flow channel and the pairing area, etc.) is exhausted, the imaging clarity of the image generation component 14 is adjusted by the focusing knob 32, and the control valves of the valve control module 20 are controlled to operate in sequence by means of the control buttons of the control panel 80 to detect whether the valve control module 20 can work normally.

[0193] After the pre-work is completed, the first light source 11a and the second light source 11b are first started, and then the valve control module 20 is used to pass the cell solution in the liquid reservoir 33 into the first particle flow channel, and the microsphere solution is passed into the second particle flow channel. When a cell passes through the first particle flow channel and is irradiated by the first detection light, the cell is excited by the first detection light to emit the first excitation light and is received by the first detection device 12a. The first detection device 12a controls the first particle flow channel control valve to block the first particle flow channel. At this time, the cell solution is cut off and stops entering the first particle flow channel, and the cell solution that has entered Under the action of inertia, the cells in the first particle flow channel flow through the first particle flow channel to the pairing area and then stop moving; when a microsphere passes through the second particle flow channel and is irradiated by the second detection light, the microsphere is excited by the second detection light to emit a second excitation light and is received by the second detection device 12b. The second detection device 12b controls the second particle flow channel control valve to block the second particle flow channel. At this time, the microsphere solution is cut off and stops entering the second particle flow channel. The microspheres that have entered the second particle flow channel flow through the second particle flow channel to the pairing area under the action of inertia and then stop moving. Subsequently, the buffer solution in the liquid reservoir 33 is passed into the pairing area through the valve control module 20, so that the buffer solution drives the cells and microspheres to approach or contact each other, and then the oil in the liquid reservoir 33 is passed into the pairing area through the valve control module 20, so that the oil solution co-encapsulates the cells and microspheres, forming droplets in which a single cell is paired with a single microsphere. The droplets are discharged to the sample collection tube 35 through the sample outlet of the microfluidic chip 2.

[0194] Then, the valve control module 20 controls the first particle flow channel control valve and the second particle flow channel control valve to reset, so that the cell solution can continue to flow in the first particle flow channel and the microsphere solution can continue to flow in the second particle flow channel, so as to carry out the next round of identification and pairing of cells and microspheres.

[0195] On the third aspect, the present application also provides an optical module for identifying first and second particles, which includes an objective lens 40, a first optical system, and a second optical system, wherein the first optical system is used to form a first detection light passing through the objective lens 40, and is used to form a second detection light passing through the objective lens 40. The first particle can be irradiated by the first detection light to emit a first excitation light, and the second particle can be irradiated by the second detection light to emit a second excitation light. The second optical system is used to receive imaging light from the microfluidic chip 2 and through the objective lens 40 to form an image of the first particle and / or the second particle. In this way, it is convenient for the user to directly check whether the particle has reached the corresponding position in the flow channel.

[0196] In this embodiment, the objective lens 40 is configured to at least adjust the spot size of the first detection light and the second detection light. Considering that the structural size of the microfluidic chip 2 is small, if the spot area is large, it may cover some inappropriate positions, such as covering another particle flow channel, thereby causing detection errors. That is, the detection module 10 has high requirements for the fineness of the spot when detecting the first particle flow channel and the second particle flow channel. Under the premise of being able to cover the detection position of the corresponding particle flow channel, reducing the area of ​​the spot is conducive to reducing interference. Based on this, the so-called adjustment of the spot size in this embodiment specifically refers to reducing the spot size of the detection module 10 by setting the objective lens 40, so that the detection light of the detection module 10 can accurately irradiate the first particle flow channel and the second particle flow channel, and make the size of the detection light of the detection module 10 adapt to the size of the first particle flow channel and the second particle flow channel, thereby facilitating the detection of the first particle flow channel and the second particle flow channel by the detection module 10.

[0197] In addition, the objective lens 40 of this embodiment can also be applied to the imaging system to assist in magnifying the image. As mentioned above, the structural size of the microfluidic chip 2 is relatively small, and the first particle flow channel, the second particle flow channel, the first particle and the second particle are difficult to see clearly with the naked eye. Therefore, the imaging light of this embodiment will also pass through the objective lens 40, and the image will be magnified by the objective lens 40, so that the user no longer needs to use other tools to see the first particle flow channel, the second particle flow channel, the first particle and the second particle.

[0198] On the basis of the third aspect of the embodiment, in some embodiments of the present application, the objective lens 40 can also be used to shape the light spot. Specifically, the cross-section of the light beam emitted by the light source is usually rectangular or circular, and its coverage area is large. The shape is not suitable for fine flow channels such as the microfluidic chip 2. The objective lens 40 of this embodiment can shape the first detection light and the second detection light so that the light spot irradiated on the microfluidic chip 2 is a strip-shaped light spot (for example, Figure 13As shown), at this time, the objective lens 40 is configured to at least adjust the corresponding size of the strip light spot, for example, the objective lens 40 is configured to adjust the width of the strip light spot so that the light spot can be concentrated on the detection position. For another example, the objective lens 40 is configured to adjust the length of the strip light spot to adapt to particle flow channels of different widths.

[0199] On the basis of the third aspect embodiment, in some embodiments of the present application, referring to Figure 1 The first optical system includes a laser emitting mechanism 11 and a detection component 12. The laser emitting mechanism 11 is used to emit a first detection light to the first particle flow channel via the objective lens 40 and the detection window 31 in sequence, and can also emit a second detection light to the second particle flow channel. The first detection light is used to make the first particles in the first particle flow channel emit a first excitation light, and the second detection light is used to make the second particles in the second particle flow channel emit a second excitation light. The detection component 12 is used to receive the first excitation light and control the first particle flow channel control valve to block the first particle flow channel. The detection component 12 is also used to receive the second excitation light and control the second particle flow channel control valve to block the second particle flow channel.

[0200] In this way, by setting up the laser emitting mechanism 11, the first particle and the second particle respectively emit the first excitation light and the second excitation light under the action of the laser, so that the detection component 12 controls the operation of the first particle flow channel control valve and the second particle flow channel control valve respectively under the control of the corresponding light beam, thereby realizing independent control of the first particle flow channel and the second particle flow channel, and improving the interception accuracy of the particle pairing device 1 for a single first particle and a single second particle.

[0201] It is understandable that in other embodiments, the detection module 10 may also detect the first particles and the second particles by electrode detection or visual image recognition.

[0202] The second optical system includes an image generation component 14, which is used to receive imaging light and form an image. This component facilitates the operator's observation of the operating status of the microfluidic chip 2, improves operational convenience, and facilitates visual detection and manual identification of the first and second particles. This not only enables the particle pairing device 1 to have different detection methods for the first and second particles, thereby enriching the functionality of the particle pairing device 1, but also interlocks the functions of the detection module 10 and the valve control module 20 through visual detection or manual identification, thereby improving the reliability of the particle pairing device 1 in pairing the first and second particles and enhancing the quality of the particle pairing. The image generation component 14 may include a camera and a display screen.

[0203] In this embodiment, the first optical system and the second optical system include a first common optical path, which is roughly marked by the area surrounded by the long dashed line in the figure. The first common optical path specifically refers to the pipeline portion shared by the excitation light and the imaging light. Figure 2 、 Figure 3 The optical system includes a dichroic mirror 1301, which is an optical element capable of transmitting or reflecting light of three or more different wavelengths or wavelength ranges. In this embodiment, the dichroic mirror 1301 is configured to reflect the first detection light and the second detection light toward the objective lens 40, and transmit the first excitation light, the second excitation light, and the imaging light. This enables reflection and transmission of different light types through a single optical element, which improves the reusability and compactness of the structure. It should be noted that the dichroic mirror 1301 can utilize known techniques.

[0204] On the basis of the embodiments of the third aspect, in some embodiments of the present application, the first optical system further includes a detection component 12, and the detection component 12 can be understood with reference to the aforementioned embodiments.

[0205] Reference Figure 1 、 Figure 8 The first common optical path also includes a first dichroic mirror 15. The first dichroic mirror 15 is arranged on the optical path between the multi-color mirror 1301 and the detection component 12. The first dichroic mirror 15 is configured to: reflect the imaging light emitted after passing through the multi-color mirror 1301 to the image generation component 14, and transmit the first excitation light and the second excitation light emitted after passing through the multi-color mirror 1301 to the detection component 12. In this way, by providing the first dichroic mirror 15, the first excitation light and the second excitation light can pass through the first dichroic mirror 15 to facilitate subsequent entry into the detection component 12, and the imaging light of other wavelengths can be reasonably utilized to generate an image through the image generation component 14, so as to facilitate observation of the working status of the microfluidic chip 2. It should be noted that the first dichroic mirror 15 can adopt a well-known technology.

[0206] As an alternative to the aforementioned embodiment, the first dichroic mirror 15 is configured to transmit the imaging light that passes through the multichromatic mirror 1301 and is emitted to the image generation component 14, and reflect the first excitation light and the second excitation light that pass through the multichromatic mirror 1301 and are emitted to the detection component 12. That is, the difference between this embodiment and the aforementioned embodiment is that the relationship between transmission and reflection is opposite.

[0207] On the basis of the third aspect embodiment, in some embodiments of the present application, referring to Figure 1 、 Figure 7The detection component 12 includes a first detection device 12a and a second detection device 12b. The first detection device 12a is electrically connected to the first particle flow channel control valve. The first detection device 12a is used to receive the first excitation light and control the first particle flow channel control valve to block the first particle flow channel. The second detection device 12b is electrically connected to the second particle flow channel control valve. The second detection device 12b is used to receive the second excitation light and control the second particle flow channel control valve to block the second particle flow channel.

[0208] In this embodiment, refer to Figure 1 、 Figure 7 The first optical system also includes a second dichroic mirror 1311, which is arranged on the optical path between the multichromatic mirror 1301 and the detection component 12. The second dichroic mirror 1311 is configured to: reflect the first excitation light that passes through the multichromatic mirror 1301 and is emitted to the first detection device 12a, and transmit the second excitation light that passes through the multichromatic mirror 1301 and is emitted to the second detection device 12b.

[0209] Specifically, the second dichroic mirror 1311 is used to change the optical paths of the first excitation light and the second excitation light, so that the first excitation light enters the first detection device 12a through the objective lens 40, the multichromatic mirror 1301 and the second dichroic mirror 1311 in sequence, and the second excitation light enters the second detection device 12b through the objective lens 40, the multichromatic mirror 1301 and the second dichroic mirror 1311 in sequence.

[0210] In this way, the first excitation light and the second excitation light have different transmission directions through the second dichroic mirror 1311, so that the two light beams can enter the first detection device 12a and the second detection device 12b respectively, thereby controlling the operation of the first particle flow channel control valve and the second particle flow channel control valve respectively, reducing the signal interference between the two light beams, avoiding signal crossing, and thereby improving the control accuracy of the first particle flow channel and the second particle flow channel.

[0211] The embodiments of the present application have been described in detail above with reference to the accompanying drawings. However, the present application is not limited to the above embodiments. Various modifications can be made within the scope of knowledge possessed by ordinary technicians in the relevant technical field without departing from the purpose of the present application. In addition, the embodiments of the present application and the features of the embodiments can be combined with each other unless there is a conflict.

Claims

1. A particle pairing device, characterized in that: Used to control the pairing of a first particle and a second particle in a microfluidic chip, the microfluidic chip is provided with a pairing area and a first particle flow channel and a second particle flow channel communicating with the pairing area, the particle pairing device comprising: a detection module, the detection module being configured to identify first particles when the first particle solution flows through the first particle flow channel, and the detection module being configured to identify second particles when the second particle solution flows through the second particle flow channel; A valve control module is electrically connected to the detection module, and is used to block the first particle flow channel when the detection module identifies a first particle, and to block the second particle flow channel when the detection module identifies a second particle.

2. The particle pairing device according to claim 1, wherein The detection module includes a first optical system, and the particle pairing device also includes an objective lens. The first optical system is used to form a first detection light that passes through the objective lens and illuminates the first particle flow channel, and is used to form a second detection light that passes through the objective lens and illuminates the second particle flow channel. The first particles can be illuminated by the first detection light to emit a first excitation light, and the second particles can be illuminated by the second detection light to emit a second excitation light, wherein the objective lens is configured to at least adjust the spot size of the first detection light and the second detection light.

3. The particle pairing device according to claim 2, characterized in that The light spot is a stripe-shaped light spot, and the objective lens is configured to at least adjust the width and / or length of the stripe-shaped light spot of the first detection light and the second detection light.

4. The particle pairing device according to claim 2, wherein: The detection module includes a second optical system, which is configured to receive imaging light from the microfluidic chip and pass through the objective lens to form an image of the microfluidic chip, wherein the objective lens is further configured to magnify the image.

5. The particle pairing device according to claim 4, characterized in that: The objective lens is configured such that a direction in which the first detection light and the second detection light pass through the objective lens is opposite to a direction in which the imaging light passes through the objective lens.

6. The particle pairing device according to claim 4, characterized in that The first optical system includes a first light source and a second light source, the first light source is used to emit the first detection light, the second light source is used to emit the second detection light, and the second optical system includes an image generating component, the image generating component is used to receive the imaging light and form the image; The first optical system and the second optical system include a first common optical path, the first common optical path includes a dichroic mirror, and the dichroic mirror is configured to: reflect the first detection light and the second detection light to the objective lens, and transmit the first excitation light, the second excitation light and the imaging light.

7. The particle pairing device according to claim 6, characterized in that The first optical system further includes a detection component configured to receive the first excitation light and the second excitation light, and identify the first particle based on the first excitation light, and identify the second particle based on the second excitation light; The first common optical path further includes a first dichroic mirror, which is disposed on the optical path between the dichroic mirror and the detection component, and is configured to: reflect the imaging light emitted from the dichroic mirror to the image generation component, and transmit the first excitation light and the second excitation light emitted from the dichroic mirror to the detection component; Alternatively, the first dichroic mirror is configured to transmit the imaging light emitted through the multichromatic mirror to the image generating component, and reflect the first excitation light and the second excitation light emitted through the multichromatic mirror to the detecting component.

8. The particle pairing device according to claim 6, wherein: The first optical system further includes a detection assembly, the detection assembly including a first detection device and a second detection device, the first detection device being configured to receive the first excitation light and identify the first particles based on the first excitation light, and the second detection device being configured to receive the second excitation light and identify the second particles based on the second excitation light; In which, the first optical system also includes a second dichroic mirror, which is arranged on the optical path between the polychroic mirror and the detection component, and the second dichroic mirror is configured to: reflect the first excitation light that passes through the polychroic mirror and is emitted to the first detection device, and transmit the second excitation light that passes through the polychroic mirror and is emitted to the second detection device.

9. The particle pairing device according to claim 6, characterized in that: The first common optical path also includes a first reflector, which is arranged on the optical path between the multichromatic mirror and the image generating component, and is configured to reflect the first excitation light, the second excitation light and the imaging light that pass through the multichromatic mirror and are emitted.

10. The particle pairing device according to claim 9, characterized in that: The objective lens, the multichromatic mirror and the first reflector are arranged in a vertical direction, and the first reflector is configured so that the first excitation light, the second excitation light and the imaging light that pass through the multichromatic mirror and are emitted are emitted toward the first reflector in a vertical direction, and are emitted in a horizontal direction after being reflected by the first reflector.

11. The particle pairing device according to claim 4, characterized in that The second optical system further includes an image generating component and a quarter wave plate. The image generating component is used to receive the imaging light and form the image. The quarter wave plate is arranged on the optical path between the objective lens and the image generating component.

12. The particle pairing device according to claim 11, characterized in that: The first optical system includes a first light source and a second light source, the first light source is used to emit the first detection light, and the second light source is used to emit the second detection light; The second optical system further includes a dichroic mirror, which is disposed on an optical path between the objective lens and the image generating assembly, and is configured to: reflect the first detection light and the second detection light to the objective lens, and transmit the first excitation light, the second excitation light, and the imaging light; Wherein, the first optical system further includes a polarization beam splitter cube, and the polarization beam splitter cube is arranged on the optical path between the first light source, the second light source and the dichroic mirror.

13. The particle pairing device according to claim 2, characterized in that The first optical system includes a first light source, a second light source and a dichroic mirror, the first light source is used to emit the first detection light, the second light source is used to emit the second detection light, and the dichroic mirror is arranged on the optical path between the first light source, the second light source and the objective lens, wherein the dichroic mirror is configured to: reflect the first detection light and the second detection light to the objective lens, and transmit the first particle to emit the first excitation light after being irradiated by the first detection light, and the second particle to emit the second excitation light after being irradiated by the second detection light.

14. The particle pairing device according to claim 13, wherein: The first optical system further includes a third dichroic mirror, which is disposed on an optical path between the first light source, the second light source and the multichroic mirror. The third dichroic mirror is configured to reflect the first detection light and transmit the second detection light.

15. The particle pairing device according to claim 14, characterized in that The first optical system further includes a second reflecting mirror, the second reflecting mirror being disposed on an optical path between the second light source and the third dichroic mirror, the second reflecting mirror being configured to reflect the second detection light toward the third dichroic mirror; The optical axis of the first light source is parallel to the optical axis of the second light source, and the first light source and the second light source are distributed in a horizontal direction.

16. The particle pairing device according to claim 2, characterized in that The first detection light and the second detection light have different wavelengths.

17. The particle pairing device according to claim 2, characterized in that In a direction from the first particle flow channel to the pairing area, the first detection light is perpendicular to the first particle flow channel, and in a direction from the second particle flow channel to the pairing area, the second detection light is perpendicular to the second particle flow channel.

18. The particle pairing device according to claim 2, characterized in that: The first detection light and the second detection light enter the objective lens at different incident positions, and / or the first detection light and the second detection light enter the objective lens at different incident angles.

19. The particle pairing device according to any one of claims 2 to 18, characterized in that: The particle pairing device further includes a supporting platform, which is used to mount the microfluidic chip. The objective lens is arranged on the other side of the supporting platform that is different from the microfluidic chip.

20. The particle pairing device according to claim 19, wherein: The support platform is provided with a detection window corresponding to the microfluidic chip, and the detection window is connected to two opposite sides of the support platform. Along the height direction of the particle pairing device, the projections of the pairing area, the first particle flow channel and the second particle flow channel are all located within the projection of the detection window, so that the objective lens can observe the microfluidic chip through the detection window.

21. The particle pairing device according to claim 19, wherein The particle pairing device further includes a three-axis motion mechanism, the support platform is mounted on the three-axis motion mechanism, and the three-axis motion mechanism is used to drive the support platform to move along three directions perpendicular to each other.

22. The particle pairing device according to claim 1, wherein The detection module includes a second optical system, and the particle pairing device also includes an objective lens. The second optical system is configured to receive imaging light from the microfluidic chip and pass through the objective lens to form an image of the microfluidic chip, wherein the objective lens is further configured to magnify the image.

23. The particle pairing device according to any one of claims 1 to 18, characterized in that The particle pairing device is provided with a liquid reservoir, which stores a first particle solution, a second particle solution, a buffer solution and an oil. The liquid reservoir is used to introduce the first particle solution into the first particle flow channel, introduce the second particle solution into the second particle flow channel, and introduce the buffer solution and the oil into the pairing area.

24. The particle pairing device according to any one of claims 1 to 18, characterized in that The particle pairing device is provided with a plurality of sample collection slots and a plurality of sample collection tubes. The plurality of sample collection slots correspond one-to-one to the plurality of sample collection tubes, and the sample collection tubes are placed in the sample collection slots. The sample collection tubes are used to collect droplets generated by the microfluidic chip.

25. The particle pairing device according to any one of claims 1 to 18, characterized in that The particle pairing device further includes a control panel, which is electrically connected to the valve control module and is used to control the start and stop of the valve control module.

26. The particle pairing device according to any one of claims 1 to 18, characterized in that The valve control module includes at least a first particle flow channel control valve and a second particle flow channel control valve. The first particle flow channel control valve is used to block the first particle flow channel when the detection module identifies a first particle, and the second particle flow channel control valve is used to block the second particle flow channel when the detection module identifies a second particle.

27. A particle pairing system, characterized in that: It comprises a microfluidic chip and a particle pairing device according to any one of claims 1 to 26, wherein the microfluidic chip is provided with a pairing area and a first particle flow channel and a second particle flow channel connected to the pairing area, and the microfluidic chip is installed in the particle pairing device.

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