Particle pairing system

Abstract

The application discloses a particle pairing device, a pairing system and an optical module. The pairing device is used for controlling pairing of a first particle and a second particle in a microfluidic chip. 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 the first particle when a first particle solution flows through a first particle flow channel of the microfluidic chip. The detection module is also used for identifying the second particle when a second particle solution flows through a second particle flow channel of the microfluidic chip. The detection module 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 identifies the first particle, and is used for blocking the second particle flow channel when the detection module identifies the second particle. Thus, 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 cells and microspheres respectively, the accuracy and efficiency of pairing of the cells and the microspheres can be improved, and thus the efficiency of single-cell sequencing can be improved.

Description

Technical Field

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

[0002] Single-cell RNA sequencing (scRNA-seq) technology reveals the heterogeneity of cells within tissues. Single-cell sequencing is widely used due to its advantages of high accuracy and high resolution compared to traditional gene sequencing technologies, providing information at the single-cell level. Single-cell sequencing technology captures genetic information based on a device that co-encapsulates cells and coding microspheres. One existing technology uses microvalve to trap cells or microspheres and uses fluid in the cell and microsphere channels to push them out, causing them to be encapsulated in oil and form droplets. During this process, multiple microspheres or cells may be encapsulated, and this technology is prone to causing contamination by empty droplets, affecting subsequent sequencing data. Furthermore, external light beams illuminating the microfluidic chip easily cause reflections, resulting in a cluttered and unclear image observed by the operator during the pairing process. This prevents the operator from accurately observing and identifying cells and microspheres during the pairing operation, further affecting the quality of cell-microsphere pairing.

[0003] Application content

[0004] 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 and a single second particle to ensure the quality of particle pairing. For example, when the first particle and the second particle are a cell and a microsphere, respectively, it can improve the accuracy and efficiency of cell-microsphere pairing, thereby improving the efficiency of single-cell sequencing.

[0005] This application also proposes a particle pairing system and an optical module.

[0006] A particle pairing device according to a first embodiment of this application is used to control the pairing of a first particle and a second particle in a microfluidic chip. The microfluidic chip has a pairing region and a first particle flow channel and a second particle flow channel communicating with the pairing region. The particle pairing device includes:

[0007] The detection module is used to identify a first particle when the first particulate solution flows through the first particulate channel, and the detection module is also used to identify a second particle when the second particulate solution flows through the second particulate channel;

[0008] A valve control module is electrically connected to the detection module. The valve control module is used to block the flow channel of the first particle when the detection module detects the first particle, and to block the flow channel of the second particle when the detection module detects the second particle.

[0009] The particle pairing device according to the embodiments of this application has at least the following beneficial effects:

[0010] This particle pairing device includes a detection module and a valve control module. The detection module is electrically connected to the valve control module, allowing the detection module to identify the first particle as it flows through the first particle channel. Simultaneously, the valve control module blocks the first particle channel to control the number of first particles within it. Similarly, the detection module identifies the second particle as it flows through the second particle channel, and the valve control module blocks the second particle channel to control the number of second particles within it. This allows for better control of the number of first and second particles entering the pairing area during the pairing process, enabling the pairing of a single first particle and a single second particle within the microfluidic chip. This ensures that a single droplet contains only a single first particle and a single second particle, guaranteeing the quality of particle pairing. For example, when the first and second particles are a cell and a microsphere, respectively, this improves the accuracy and efficiency of cell-microsphere pairing, reducing time and material waste caused by unqualified paired samples, and ultimately improving the efficiency of single-cell sequencing.

[0011] In other embodiments of this application, the detection module includes a first optical system, and the particle pairing device further includes an objective lens. The first optical system is configured to form a first detection light passing through the objective lens and illuminating the first particle channel, and to form a second detection light passing through the objective lens and illuminating the second particle channel. The first particle can be illuminated by the first detection light to emit a first excitation light, and the second particle can be illuminated by the second detection light to emit a second excitation light. The objective lens is configured to at least adjust the spot size of the first detection light and the second detection light.

[0012] In other embodiments of this 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.

[0013] In other embodiments of this application, the detection module includes a second optical system 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.

[0014] In other embodiments of this application, the objective lens is configured such that the first detection light and the second detection light pass through the objective lens in opposite directions, and the imaging light passes through the objective lens in opposite directions.

[0015] In other embodiments of this application, the first optical system includes a first light source and a second light source, the first light source being used to emit the first detection light, the second light source being used to emit the second detection light, and the second optical system including an image generation component being used to receive the imaging light and form the image;

[0016] The first optical system and the second optical system include a first common optical path, which includes a multi-directional color mirror. The multi-directional color 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.

[0017] In other embodiments of this application, the first optical system further includes a detection component, which is configured to receive the first excitation light and the second excitation light, and to identify the first particle based on the first excitation light, and to identify the second particle based on the second excitation light;

[0018] The first common optical path further includes a first dichroic mirror, which is disposed on the optical path between the multi-directional mirror and the detection component. The first dichroic mirror is configured to reflect the imaging light emitted from the multi-directional mirror to the image generation component, and to transmit the first excitation light and the second excitation light emitted from the multi-directional mirror to the detection component.

[0019] Alternatively, the first dichroic mirror is configured to transmit the imaging light emitted after passing through the dichroic mirror to the image generation component, and to reflect the first excitation light and the second excitation light emitted after passing through the dichroic mirror to the detection component.

[0020] In other embodiments of this application, the first optical system further includes a detection component, which includes a first detection device and a second detection device. The first detection device is used to receive the first excitation light and identify the first particle based on the first excitation light. The second detection device is used to receive the second excitation light and identify the second particle based on the second excitation light.

[0021] The first optical system further includes a second dichroic mirror, which is disposed in the optical path between the multi-directional mirror and the detection component. The second dichroic mirror is configured to reflect the first excitation light emitted after passing through the multi-directional mirror to the first detection device, and to transmit the second excitation light emitted after passing through the multi-directional mirror to the second detection device.

[0022] In other embodiments of this application, the first common optical path further includes a first reflector disposed on the optical path between the multi-directional color mirror and the image generation component. The first reflector is configured to reflect the first excitation light, the second excitation light, and the imaging light emitted after passing through the multi-directional color mirror.

[0023] In other embodiments of this application, the objective lens, the multi-color mirror, and the first reflecting mirror are arranged vertically. The first reflecting mirror is configured such that the first excitation light, the second excitation light, and the imaging light emitted after passing through the multi-color mirror are directed vertically toward the first reflecting mirror and emitted horizontally after being reflected by the first reflecting mirror.

[0024] In other embodiments of this application, the second optical system further includes an image generating component and a quarter-wave plate, the image generating component being used to receive the imaging light and form the image, and the quarter-wave plate being disposed in the optical path between the objective lens and the image generating component.

[0025] In other embodiments of this application, the first optical system includes a first light source and a second light source, wherein 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;

[0026] The second optical system further includes a multi-directional color mirror disposed in the optical path between the objective lens and the image generating component. The multi-directional color 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.

[0027] The first optical system further includes a polarization beam splitter cube, which is disposed in the optical path between the first light source, the second light source and the multi-directional color mirror.

[0028] In other embodiments of this application, the first optical system includes a first light source, a second light source, and a multi-directional 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 multi-directional mirror is disposed in the optical path between the first light source, the second light source, and the objective lens. The multi-directional mirror is configured to reflect the first detection light and the second detection light to the objective lens, and transmit the first particle emitting a first excitation light after being irradiated by the first detection light, and the second particle emitting a second excitation light after being irradiated by the second detection light.

[0029] In other embodiments of this application, the first optical system further includes a third dichroic mirror, which is disposed in the optical path between the first light source, the second light source and the dichroic mirror, and is configured to reflect the first detection light and transmit the second detection light.

[0030] In other embodiments of this application, the first optical system further includes a second reflector disposed in the optical path between the second light source and the third dichroic mirror, and the second reflector is configured to reflect the second detection light to the third dichroic mirror;

[0031] 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.

[0032] In other embodiments of this application, the wavelengths of the first detection light and the second detection light are different.

[0033] In other embodiments of this application, the first detection light is perpendicular to the first particle channel in the direction from the first particle channel to the pairing region, and the second detection light is perpendicular to the second particle channel in the direction from the second particle channel to the pairing region.

[0034] In other embodiments of this 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.

[0035] In other embodiments of this application, the particle pairing device further includes a support platform for mounting the microfluidic chip, and the objective lens is disposed on the other side of the support platform, distinct from the microfluidic chip.

[0036] In other embodiments of this application, the support platform is provided with a detection window corresponding to the microfluidic chip. The detection window connects the 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 channel, and the second particle 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.

[0037] In other embodiments of this application, the particle pairing device further includes a three-axis motion mechanism, on which the support platform is mounted, and the three-axis motion mechanism is used to drive the support platform to move in three mutually perpendicular directions.

[0038] In other embodiments of this application, the detection module includes a second optical system, and the particle pairing device further 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.

[0039] In other embodiments of this application, the particle pairing device is provided with a storage tank, which stores a first particle solution, a second particle solution, a buffer solution, and an oil. The storage tank is used to introduce the first particle solution into the first particle channel, introduce the second particle solution into the second particle channel, and introduce the buffer solution and oil into the pairing area.

[0040] In other embodiments of this application, the particle pairing device is provided with a plurality of sample collection slots and a plurality of sample collection tubes, wherein the plurality of sample collection slots correspond one-to-one with the plurality of sample collection tubes, and the sample collection tubes are placed in the sample collection slots, and the sample collection tubes are used to collect droplets generated by the microfluidic chip.

[0041] In other embodiments of this application, the particle pairing device further includes a control panel electrically connected to the valve control module, the control panel being used to control the start and stop of the valve control module.

[0042] In other embodiments of this 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 detects a first particle, and the second particle flow channel control valve is used to block the second particle flow channel when the detection module detects a second particle.

[0043] The particle pairing system according to the second embodiment of this application includes a microfluidic chip and the particle pairing device. The microfluidic chip has a pairing area and a first particle channel and a second particle channel communicating with the pairing area, and the microfluidic chip is installed in the particle pairing device.

[0044] An optical module according to a third embodiment of this application is used to identify a first particle and a second particle, including:

[0045] Objective lens;

[0046] A first optical system is configured to generate a first detection light passing through the objective lens and a second detection light passing through the objective lens, wherein 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.

[0047] A second optical system is used to receive imaging light from the microfluidic chip and through the objective lens to form images of the first particle and / or the second particle;

[0048] The objective lens is configured to adjust the spot size of the first detection light and the second detection light, and to magnify the image.

[0049] In other embodiments of this 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.

[0050] In other embodiments of this application, the first optical system includes a first light source and a second light source, the first light source being used to emit the first detection light, the second light source being used to emit the second detection light, and the second optical system including an image generation component being used to receive the imaging light and form the image;

[0051] The first optical system and the second optical system include a first common optical path, which includes a multi-directional color mirror. The multi-directional color 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.

[0052] In other embodiments of this application, the first optical system further includes a detection component, which is configured to receive the first excitation light and the second excitation light, and to identify the first particle based on the first excitation light, and to identify the second particle based on the second excitation light;

[0053] The first common optical path further includes a first dichroic mirror, which is disposed on the optical path between the multi-directional mirror and the detection component. The first dichroic mirror is configured to reflect the imaging light emitted from the multi-directional mirror to the image generation component, and to transmit the first excitation light and the second excitation light emitted from the multi-directional mirror to the detection component.

[0054] Alternatively, the first dichroic mirror is configured to transmit the imaging light emitted after passing through the dichroic mirror to the image generation component, and to reflect the first excitation light and the second excitation light emitted after passing through the dichroic mirror to the detection component.

[0055] In other embodiments of this application, the first optical system further includes a detection component, which includes a first detection device and a second detection device. The first detection device is used to receive the first excitation light and identify the first particle based on the first excitation light. The second detection device is used to receive the second excitation light and identify the second particle based on the second excitation light.

[0056] The first optical system further includes a second dichroic mirror, which is disposed in the optical path between the multi-directional mirror and the detection component. The second dichroic mirror is configured to reflect the first excitation light emitted after passing through the multi-directional mirror to the first detection device, and to transmit the second excitation light emitted after passing through the multi-directional mirror to the second detection device.

[0057] Additional aspects and advantages of this application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of this application. Attached Figure Description

[0058] The present application will be further described below with reference to the accompanying drawings and embodiments, wherein:

[0059] Figure 1 This is a schematic diagram of the particle pairing device disclosed in the embodiments of this application from one perspective;

[0060] Figure 2 This is a schematic diagram of the particle pairing device disclosed in the embodiments of this application from another perspective;

[0061] Figure 3 This is a right view of the particle pairing device disclosed in the embodiments of this application;

[0062] Figure 4 This is a schematic diagram of the optical path of the main display objective lens of the particle pairing device disclosed in the embodiments of this application;

[0063] Figure 5 This is a schematic diagram of the optical path of the particle pairing device mainly displaying the detection component, as disclosed in the embodiments of this application.

[0064] Figure 6 This is a schematic diagram of the optical path of the main display image generation component of the particle pairing device disclosed in the embodiments of this application;

[0065] Figure 7 This is a schematic diagram of the optical path showing the specific composition of the detection components of the particle pairing device disclosed in the embodiments of this application;

[0066] Figure 8 This is a schematic diagram of the optical path of the particle pairing device disclosed in the embodiments of this application, mainly showing the first common optical path;

[0067] Figure 9 This is a schematic diagram of the optical path of the first reflecting mirror of the particle pairing device disclosed in the embodiments of this application;

[0068] Figure 10 This is a schematic diagram of the optical path of the particle pairing device disclosed in the embodiments of this application, mainly showing the anti-reflection structure;

[0069] Figure 11 This is a schematic diagram of the optical path of the particle pairing device disclosed in the embodiments of this application, mainly showing the specific structure of the laser emitting mechanism;

[0070] Figure 12 This is a schematic diagram of the optical path of the particle pairing device disclosed in the embodiments of this application;

[0071] Figure 13 This is a schematic diagram showing the first laser and the second laser disclosed in the embodiments of this application being perpendicular to the first particle flow channel and the second particle flow channel, respectively;

[0072] Figure 14 This is a schematic diagram of the microfluidic chip to which this application applies;

[0073] Figure 15 This is a schematic diagram of the particle pairing device disclosed in an embodiment of this application from one perspective. The support platform, three-axis motion mechanism and control panel are hidden in the figure.

[0074] Figure 16 for Figure 15 Top view;

[0075] Figure 17 This is a schematic diagram showing the connection between the support platform and the three-axis motion mechanism disclosed in the embodiments of this application.

[0076] Figure label:

[0077] Particle pairing device 1;

[0078] Microfluidic chip 2, first particle channel 21, second particle channel 22, oil channel 23, buffer channel 24, waste liquid channel 25, paired channel 26, first micro valve 28, second micro valve 29, buffer micro valve 210, waste liquid micro valve 211, oil micro valve 212;

[0079] Detection module 10, laser emitting mechanism 11, first light source 11a, second light source 11b, detection component 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 component 14, first dichroic mirror 15;

[0080] Valve control module 20;

[0081] Support platform 30, detection window 31, focusing knob 32, liquid storage tank 33, sample collection tank 34, sample collection tube 35, fixture 36;

[0082] Objective lens 40;

[0083] 50 bright field lights, 51 light poles, 52 bright field light sources;

[0084] Anti-reflection structure 60, polarization beam splitter cube 61, quarter-wave plate 62;

[0085] Three-axis motion mechanism 70, first horizontal drive component 71, second horizontal drive assembly 72, vertical drive assembly 73;

[0086] Control Panel 80;

[0087] Base 90, connecting seat 91, column 92. Detailed Implementation

[0088] The embodiments of this application are described in detail below. Examples of these embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain this application, and should not be construed as limiting this application.

[0089] In the description of this application, it should be understood that the orientation descriptions, such as up, down, front, back, left, right, etc., indicate the orientation or positional relationship based on the orientation or positional relationship 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, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this application.

[0090] In the description of this application, "several" means one or more, "multiple" means two or more, "greater than," "less than," and "exceeding" are understood to exclude the stated number, while "above," "below," and "within" are understood to include the stated number. The use of "first" and "second" in the description is merely for distinguishing technical features and should not be construed as indicating or implying relative importance, or implicitly indicating the number of indicated technical features, or implicitly indicating the order of the indicated technical features.

[0091] In the description of this application, unless otherwise expressly defined, terms such as "setup," "installation," and "connection" should be interpreted broadly, and those skilled in the art can reasonably determine the specific meaning of the above terms in this application in conjunction with the specific content of the technical solution.

[0092] In the description of this application, the terms "one embodiment," "some embodiments," "illustrative embodiment," "example," "specific example," or "some examples," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0093] The technical solution of this application will be further described below with reference to the embodiments and accompanying drawings.

[0094] Please refer to the following: Figures 1 to 4 An embodiment of the first aspect of this 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 has a pairing region and a first particle flow channel and a second particle flow channel connecting 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 used to identify the first particle when the first particle solution flows through the first particle flow channel, and the detection module 10 is also used to identify the second particle when the second particle solution flows through the second particle flow channel. The detection module 10 is electrically connected to the valve control module 20, and the valve control module 20 is used 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.

[0095] This application provides a particle pairing device 1, including a detection module 10 and a valve control module 20. The detection module 10 is electrically connected to the valve control module 20, allowing the detection module 10 to identify first particles as the first particle solution flows through the first particle channel. Simultaneously, the valve control module 20 blocks the first particle channel to control the number of first particles within it. Similarly, the detection module 10 identifies second particles as the second particle solution flows through the second particle channel, and the valve control module 20 blocks the second particle channel to control the number of second particles within it. This facilitates control over the number of first and second particles entering the pairing area during the pairing process, enabling the pairing of a single first particle and a single second particle in the microfluidic chip 2. That is, a single droplet contains only a single first particle and a single second particle, ensuring the quality of particle pairing. For example, when the first particle and the second particle are a cell and a microsphere, respectively, this improves the accuracy and efficiency of cell-microsphere pairing, reducing time and material costs wasted due to unqualified paired samples, thereby improving the efficiency of single-cell sequencing.

[0096] Specifically, refer to Figure 14 The diagram shows a typical microfluidic chip 2 from the front, which has multiple flow channels internally, including, for example, 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 paired flow channel 26. The first particle flow channel 21 is for a first particle solution containing first particles to pass through; the second particle flow channel 22 is for a second particle solution containing second particles to pass through; the buffer flow channel 24 is for a buffer solution to pass through; the oil flow channel 23 is for an oil solution that is immiscible with the first particle solution, the second particle solution, and the buffer solution to pass through; the waste liquid flow channel 25 is for discharging the first particle solution and the second particle solution; and the paired flow channel 26 is for a single first particle and a single second particle to remain. The paired 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 paired flow channel 26.

[0097] The microfluidic chip 2 also includes multiple microvalves, specifically including a paired microvalves 27 corresponding to the paired flow channel 26, a first microvalves 28 corresponding to the first particle flow channel 21, a second microvalves 29 corresponding to the second particle flow channel 22, a buffer microvalves 210 corresponding to the buffer flow channel 24, a waste liquid microvalves 211 corresponding to the waste liquid flow channel 25, and an oil microvalves 212 corresponding to the oil flow channel 23. The paired microvalves 27 are used to control the opening and closing of the paired flow channel 26, the first microvalves 28 are used to control the opening and closing of the first particle flow channel 21, the second microvalves 29 are used to control the opening and closing of the second particle flow channel 22, the buffer microvalves 210 are used to control the opening and closing of the buffer flow channel 24, the waste liquid microvalves 211 are used to control the opening and closing of the waste liquid flow channel 25, and the oil microvalves 212 are used to control the opening and closing of the oil flow channel 23.

[0098] Based on the above structure, the pairing process of microfluidic chip 2 is as follows:

[0099] Buffer microvalve 210 and paired microvalve 27 are closed, while first microvalve 28, second microvalve 29, and waste liquid microvalve 211 are opened, allowing the first and second particle solutions to drain through paired flow channel 26 and waste liquid flow channel 25. Upon detection of a single first particle, first microvalve 28 closes, stopping the single first particle within paired flow channel 26. Upon detection of a single second particle, second microvalve 29 closes, stopping the single second particle within paired flow channel 26. Once both the single first and single second particles have stopped within paired flow channel 26, waste liquid microvalve 211 closes.

[0100] Based on the above structure, the packaging process of microfluidic chip 2 is as follows:

[0101] When the first microvalve 28, the second microvalve 29, and the waste liquid microvalve 211 are closed, the buffer microvalve 210 and the paired microvalve 27 are opened, and the buffer liquid sends the single first particle and the single second particle in the paired flow channel 26 into the oil in the oil flow channel 23 to form droplets.

[0102] Based on the first aspect of the embodiment, the first particle and the second particle may optionally be any one of cells, microspheres or other fine particles, and the specific configuration can be set according to actual needs, without limitation here.

[0103] Based on the first aspect embodiment, 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.

[0104] In this way, by controlling the first particle flow channel and the second particle flow channel separately through multiple control valves, the blocking or connection to the outside of the first particle flow channel and the second particle flow channel can be controlled independently. This allows the corresponding flow channel to be blocked when the first particle or the second particle passes through, making the control of each flow channel by the valve control module 20 more flexible, which helps to ensure the pairing of a single first particle with a single second particle.

[0105] Based on the first aspect embodiment, optionally, both the first particle flow channel control valve and the second particle flow channel control valve can be any one of pneumatic valves, electric valves, or hydraulic valves; or, each can be a pneumatic valve, electric valve, or hydraulic valve. For example, both the first and second particle flow channel control valves are pneumatic valves, thereby making the control method of the valve control module 20 for the first and second particle flow channels simple, safe, and reliable, thus improving the reliability of the pairing of the first and second particles and ensuring the quality of particle pairing.

[0106] Based on the first aspect of the embodiments, in some embodiments of this application, the detection module 10 includes a first optical system, which is used to form a first detection light irradiating a first particle channel and a second detection light irradiating a second particle 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 fluorescence signal is captured.

[0107] In this embodiment, refer to Figures 1 to 4 The particle pairing device 1 also includes an objective lens 40, through which both the first and second detection lights pass. The objective lens 40 is configured to at least adjust the spot size of the first and second detection lights. Considering the small size of the microfluidic chip 2, if the spot area is large, it may cover some unsuitable positions, such as another particle channel, leading to detection errors. That is, the detection module 10 has high requirements for the precision of the spot when detecting the first and second particle channels. While ensuring that the detection position of the corresponding particle channel is covered, reducing the area of ​​the spot helps to reduce interference. Based on this, the adjustment of the spot size mentioned in this embodiment specifically refers to reducing the size of the spot of the detection module 10 by setting the objective lens 40, so that the detection light of the detection module 10 can accurately illuminate the first and second particle channels, and make the size of the detection light of the detection module 10 adapt to the size of the first and second particle channels, thereby facilitating the detection of the first and second particle channels by the detection module 10.

[0108] In some specific embodiments, the microfluidic chip 2 and the detection module 10 are respectively disposed on the object side (e.g., the upper side in the figure) and the image side (e.g., the lower side in the figure) of the objective lens 40.

[0109] When the detection module 10 includes a first optical system, and the first optical system includes an objective lens 40, in some embodiments of this application, the objective lens 40 can also be used for shaping the light spot. Specifically, the cross-section of the light beam emitted by the light source is usually rectangular or circular, with a large coverage area, and its shape is not suitable for the fine flow channels such as the microfluidic chip 2. In this embodiment, the objective lens 40 can shape the first detection light and the second detection light so that the light spot illuminating the microfluidic chip 2 is a strip-shaped light spot (e.g., Figure 13 As shown), at this time, the objective lens 40 is configured to at least adjust the corresponding size of the strip-shaped light spot. For example, the objective lens 40 is configured to adjust the width of the strip-shaped light spot so that the light spot can be concentrated at the detection position. Or, for example, the objective lens 40 is configured to adjust the length of the strip-shaped light spot so as to adapt to particle channels of different widths.

[0110] Meanwhile, by setting the objective lens 40 to magnify the structure of the microfluidic chip 2, it is also convenient for operators to observe with their naked eyes, so as to realize different detection methods of the second particle and the first particle, and it is also beneficial to check the function of the valve control module 20 and the detection module 10, thereby improving the functional reliability of the particle pairing device 1.

[0111] Based on the first aspect of the embodiments, optionally, refer to Figure 1 The particle pairing device 1 also includes a support platform 30, which is disposed on the object side of the objective lens 40, and 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 in a stable and stationary state when the particle pairing device 1 is working, thereby improving the working reliability of the particle pairing device 1.

[0112] Based on the first aspect of the embodiment, optionally, the support platform 30 is provided with a clamp 36 for holding the microfluidic chip 2. This facilitates the stable installation of the microfluidic chip 2 on the particle pairing device 1, enabling the particle pairing device 1 to detect and control the first and second particle channels. Simultaneously, the clamp 36 facilitates the disassembly and installation of the microfluidic chip 2, allowing for replacement, maintenance, and updates, thus enabling the particle pairing device 1 to be used with different microfluidic chips 2 and improving the practicality of the particle pairing device 1.

[0113] Based on the first aspect of the embodiments, optionally, refer 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 channel and the second particle 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.

[0114] In addition, by setting the detection window 31, the microfluidic chip 2 and the objective lens 40 can be located on both sides of the support platform 30, which is conducive to making reasonable use of space, improving the structural compactness, and facilitating the installation and removal of the microfluidic chip 2 while ensuring the stability of the structure.

[0115] For example, refer to Figure 1 The support platform 3 includes a flat main structure and is connected to the base 90 via the three-axis motion mechanism 70 described below. The 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 the clamp 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 rod, and other structures and is located below the detection window 31.

[0116] The foregoing embodiments describe the functions of adjusting the spot size and reshaping the spot using the objective lens 40. In other embodiments of this application, the objective lens 40 can also be applied to an imaging system to assist in image magnification. Specifically, the detection module 10 includes a second optical system configured to receive imaging light from the microfluidic chip 2 to form an image of the microfluidic chip 2. This allows users to directly check whether particles have reached the corresponding positions within the flow channels. As mentioned earlier, the microfluidic chip 2 has a small structural size, 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, in this embodiment, the imaging light also passes through the objective lens 40 to magnify the image, so that users do not need to use other tools to see the first particle flow channel, the second particle flow channel, the first particle, and the second particle.

[0117] In this embodiment, the objective lens 40 can realize functions such as adjusting the spot area, adjusting the spot shape, and magnifying the image, which is not only beneficial for the detection of particulate matter, but also makes it convenient for users to view the condition of particulate matter. In addition, it can simplify the structure of the detection module 10 and reduce costs.

[0118] When the detection module 10 further includes a second optical system, in some embodiments of this application, refer to Figure 4The objective lens 40 is configured such that the directions in which the first detection light and the second detection light pass through the objective lens 40 are opposite to the direction in which the imaging light passes through the objective lens 40. For example, if the first detection light and the second detection light pass through the objective lens 40 in an upward direction, then the imaging light passes through the objective lens 40 in a downward direction. Based on optical principles, for optical devices (such as convex lenses) that can adjust the direction of light after it enters, when light enters from a certain direction, it will converge, and when it enters from the opposite direction, it will diverge. In this embodiment, by setting the incident directions of the detection light and the imaging light, the reduction of the light spot and the magnification of the image can be achieved simultaneously through a single light source element.

[0119] When the detection module 10 includes a first optical system and a second optical system, in some embodiments, refer 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 into the first particle channel through the objective lens 40 and the detection window 31 in sequence, and can also emit a second detection light into the second particle channel. The first detection light is used to cause the first particle in the first particle channel to emit a first excitation light, and the second detection light is used to cause the second particle in the second particle channel to emit a second excitation light. The detection component 12 is used to receive the first excitation light and control the first particle channel control valve to block the first particle channel. The detection component 12 is also used to receive the second excitation light and control the second particle channel control valve to block the second particle channel.

[0120] In this way, by setting up the laser emitting mechanism 11, the first particle and the second particle emit the first excitation light and the second excitation light respectively under the action of the laser, so that the detection component 12 controls the first particle flow channel control valve and the second particle flow channel control valve respectively under the control of the corresponding 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.

[0121] Understandably, in other embodiments, the detection module 10 may also use electrode detection or visual image recognition to detect the first particle and the second particle.

[0122] Reference Figure 6The second optical system includes an image generation component 14, which receives imaging light and forms an image. This facilitates the operator's observation of the microfluidic chip 2's operating status, improving operational convenience and enabling visual detection and manual identification of the first and second particles. This allows the particle pairing device 1 to employ different detection methods for the first and second particles, enriching its functionality. Furthermore, visual detection or manual identification can interlock the functions of the detection module 10 and the valve control module 20, improving the reliability of the particle pairing device 1 in matching the first and second particles and thus enhancing the quality of particle pairing. The image generation component 14 may include a camera and a display screen.

[0123] In other embodiments, the first optical system and the second optical system include a first common optical path, the first common optical path being in Figure 8 The area roughly marked by the long dashed line is the first common optical path, which specifically refers to the section of the pipeline shared by the excitation light and the imaging light. (Refer to...) Figure 2 , Figure 3 and Figure 8 It includes a multi-color mirror 1301, which is an optical element capable of allowing or reflecting three or more different wavelengths or wavelength ranges of light. In this embodiment, the multi-color mirror 1301 is configured to reflect the first detection light and the second detection light to the objective lens 40, and transmit the first excitation light, the second excitation light, and the imaging light. Thus, by using a single optical element to achieve the reflection and transmission of different lights, the reusability of the structure is improved, further enhancing the structural compactness. It should be noted that the multi-color mirror 1301 can employ known technologies.

[0124] When the first optical system includes a laser emitting mechanism 11, optionally, refer 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 into 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 particle 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 into 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 particle in the second particle flow channel to generate a second excitation light. The wavelengths of the first detection light and the second detection light are different.

[0125] In this way, by irradiating the first and second particles with lasers, it is possible to ensure that the light has sufficient energy to activate the first and second particles and emit light beams with sufficient brightness. At the same time, by irradiating the first and second particle channels with two lasers of different wavelengths respectively, the first and second particles can be excited to emit light beams of different colors and wavelengths, so that the two photoelectric converters can receive the light beams emitted from the first and second particles respectively, which is conducive to the independent detection of the first and second particles.

[0126] 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., which can be set according to actual needs and are not limited here. However, it should be noted that the first detection light and the second detection light must be selected with different wavelengths. 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 components 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.

[0127] For example, in this embodiment, the wavelength of the first detection light is 488nm and the wavelength of the second detection light is 640nm, so as to satisfy both the design requirement that the wavelengths of the first detection light and the second detection light are different, and 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.

[0128] It is understood that in other embodiments, the first detection light and the second detection light may also be light of other wavelengths.

[0129] When the first optical system and the second optical system include a first common optical path, in some embodiments of this application, the first optical system further includes a detection component 12, which can be understood with reference to the foregoing embodiments.

[0130] Reference Figure 1 , Figure 8The first shared optical path also includes a first dichroic mirror 15, which is disposed in the optical path between the multi-polarized mirror 1301 and the detection component 12. The first dichroic mirror 15 is configured to reflect the imaging light emitted from the multi-polarized mirror 1301 to the image generation component 14, and to transmit the first excitation light and the second excitation light emitted from the multi-polarized mirror 1301 to the detection component 12. Thus, by setting the first dichroic mirror 15, the first and second excitation lights can pass through the first dichroic mirror 15 to facilitate their subsequent entry into the detection component 12, while other wavelengths of imaging light can be utilized to generate an image through the image generation component 14, facilitating the observation of the working state of the microfluidic chip 2. It should be noted that the first dichroic mirror 15 can employ known technology.

[0131] As an alternative to the aforementioned embodiment, the first dichroic mirror 15 is configured to transmit the imaging light emitted after passing through the dichroic mirror 1301 to the image generation component 14, and reflect the first excitation light and the second excitation light emitted after passing through the dichroic mirror 1301 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 reversed.

[0132] In the foregoing embodiments, optionally, the first dichroic mirror 15 may be a long-pass dichroic mirror. The two opposite surfaces of the first dichroic mirror 15 are respectively provided with a beam-splitting film and an anti-reflection film. The image generation component 14 is disposed on the side of the first dichroic mirror 15 where the beam-splitting film is provided, and the detection component 12 is disposed on the side of the first dichroic mirror 15 where the anti-reflection film is provided. 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 both the first excitation light and the second excitation light can 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 generation component 14 to generate an image.

[0133] For example, in this embodiment, the first dichroic mirror 15 is a 490nm long-pass dichroic mirror, so that light with a wavelength less than 490nm is reflected and enters the image generation 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 directed toward the detection component 12.

[0134] When the first optical system includes a detection component 12 and the first common optical path includes a multi-color mirror 1301, in some embodiments of this application, refer 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 and 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 and is used to receive the second excitation light and control the second particle flow channel control valve to block the second particle flow channel.

[0135] In this way, by receiving the first excitation light and the second excitation light respectively through the first detection device 12a and the second detection device 12b and controlling the operation of the two control valves respectively, the first excitation light and the second excitation light can be converted into two photoelectric signals by the first detection device 12a and the second detection device 12b respectively, so as to improve the accuracy of the photoelectric signals and avoid the formation of interference signals. This is beneficial to improving 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.

[0136] Optionally, the first detection device 12a and the second detection device 12b can be any one of a photomultiplier tube (PMT), an avalanche photodiode (APD), or a photodiode. The specific device can be set according to actual needs and is not limited here.

[0137] In this embodiment, refer to Figure 1 , Figure 7 , Figure 8 The first optical system further includes a second dichroic mirror 1311, which is disposed in the optical path between the multi-directional mirror 1301 and the detection component 12. The second dichroic mirror 1311 is configured to reflect the first excitation light emitted after passing through the multi-directional mirror 1301 to the first detection device 12a, and transmit the second excitation light emitted after passing through the multi-directional mirror 1301 to the second detection device 12b.

[0138] 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 sequentially through the objective lens 40, the multi-color mirror 1301 and the second dichroic mirror 1311, and the second excitation light enters the second detection device 12b sequentially through the objective lens 40, the multi-color mirror 1301 and the second dichroic mirror 1311.

[0139] In this way, the second dichroic mirror 1311 makes the first excitation light and the second excitation light have different transmission directions, so that the two 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 signal interference between the two beams, avoiding signal crossing, and thus improving the control accuracy of the first particle flow channel and the second particle flow channel.

[0140] Optionally, the second dichroic mirror 1311 can be a long-pass dichroic mirror. The two opposite surfaces of the second dichroic mirror 1311 are respectively provided with a beam-splitting film and an anti-reflection film. The first detection device 12a is disposed on the side of the second dichroic mirror 1311 with the beam-splitting film, and the second detection device 12b is disposed on the side of the second dichroic mirror 1311 with the anti-reflection film. 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, while the light beam with a wavelength lower than the cutoff wavelength is reflected by the second dichroic mirror 1311. This allows the first excitation light and the second excitation light to be transmitted in two different directions, so that the first excitation light enters the first detection device 12a and the second excitation light enters the second detection device 12b.

[0141] For example, in this embodiment, the second dichroic mirror 1311 is a 550nm long-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.

[0142] When the second optical system includes an image generation component 14 and the first common optical path includes a multi-color mirror 1301, in some embodiments of this application, refer to Figure 3 , Figure 9 The first shared optical path also includes a first reflecting mirror 1310, which is disposed in the optical path between the multi-color mirror 1301 and the image generating component 14. Specifically, the multi-color mirror 1301 is disposed between the objective lens 40 and the first reflecting mirror 1310 along the optical axis of the objective lens 40. The first reflecting mirror 1310 is configured to reflect the first excitation light, the second excitation light, and the imaging light emitted after passing through the multi-color mirror 1301. By adjusting the transmission direction of the first and second excitation lights using the first reflecting mirror 1310, it is beneficial to reduce the size of the particle pairing device 1 along the optical axis of the objective lens 40.

[0143] When the first common optical path also includes the first reflector 1310, in some embodiments of this application, refer to Figure 3 , Figure 9The objective lens 40, the multi-color mirror 1301, and the first reflecting mirror 1310 are arranged vertically, for example, from top to bottom. The first reflecting mirror 1310 is configured such that the first excitation light, the second excitation light, and the imaging light emitted after passing through the multi-color mirror 1301 are directed vertically towards the first reflecting mirror 1310, and after being reflected by the first reflecting mirror 1310, are emitted horizontally. This facilitates the placement of the microfluidic chip 2 at the top, making it easy for the user to operate, while also making full use of the space in both the vertical and horizontal directions, which helps control the volume of the particle pairing device 1.

[0144] It should be noted that when the first common optical path includes a multi-color mirror 1301, a first reflecting mirror 1310, a first dichroic mirror 15, and a second dichroic mirror 1311, it adopts the following layout: the objective lens 40 is located on the upper layer, the multi-color mirror 1301 is located on the middle layer, and the first reflecting mirror 1310, the first dichroic mirror 15, and the second dichroic mirror 1311 are located on the lower layer, and the multi-color mirror 1301, the first reflecting mirror 1310, the first dichroic mirror 15, and the second dichroic mirror 1311 are arranged sequentially along the emission direction of the excitation light.

[0145] Considering that the substrate of the microfluidic chip 2 is a transparent plate, strong reflections are easily formed when the laser shines on the microfluidic chip 2 through the detection window 31. This results in uneven light intensity and cluttered, unclear images captured by the image generation component 14, making it difficult for operators to clearly observe the working status of the microfluidic chip 2. Consequently, the number of cells and microspheres passing through the flow channel cannot be accurately identified, thus affecting the quality of cell-microsphere pairing. Based on this, when the detection module 10 also includes a second optical system, in some embodiments of this application, referring to... Figure 3 , Figure 10 The second optical system also includes an image generation component 14 and a quarter-wave plate 62. The image generation component 14 is used to receive imaging light and form an image, as can be understood with reference to the foregoing embodiments. The quarter-wave plate 62 is disposed in the optical path between the objective lens 40 and the image generation component 14. By setting the quarter-wave plate 62 to weaken the reflected light from the transparent plate, it is beneficial to reduce the stray light collected by the image generation component 14, improve the purity and clarity of the image, thereby improving the accuracy of cell and microsphere identification, and thus improving the quality of cell-microsphere pairing.

[0146] Specifically Figure 10 In the embodiment shown, when the first common optical path includes a multi-directional mirror 1301, a quarter-wave plate 62 is disposed on the optical path between the objective lens 40 and the multi-directional mirror 1301.

[0147] When the second optical system also includes a quarter-wave plate 62, in some embodiments of this application, refer 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 emits a first detection light, and the second light source 11b emits a second detection light. The second optical system also includes a multi-color mirror 1301, which is disposed in the optical path between the objective lens 40 and the image generation component 14. The multi-color mirror 1301 is configured to reflect the first and second detection lights to 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 multi-color mirror 1301 can all be understood with reference to the foregoing embodiments.

[0148] 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 multi-directional color mirror 1301. The polarization beam splitter cube 61 can work with the quarter-wave plate 62 to further reduce the reflected light from the transparent plate.

[0149] The foregoing embodiments proposed that the objective lens can achieve functions such as adjusting the spot area, adjusting the spot shape, and magnifying the image, and proposed a first optical system and a second optical system based on this. In other embodiments of this application, a scheme for the first optical system is proposed 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 multi-color mirror 1301. The first light source 11a is used to emit a first detection light, the second light source 11b is used to emit a second detection light, and the multi-color mirror 1301 is disposed in the optical path between the first light source 11a, the second light source 11b, and the objective lens 40. The multi-color mirror 1301 is configured to reflect the first and second detection lights to the objective lens 40, and transmit the first excitation light emitted by a first particle after being irradiated by the first detection light, and the second excitation light emitted by a second particle after being irradiated by the second detection light. The first light source 11a, the second light source 11b, and the multi-color mirror 1301 in this embodiment can all be understood with reference to the foregoing embodiments.

[0150] Based on the foregoing embodiments, in some embodiments of this application, reference is made to... Figure 2 , Figure 11 The first optical system also includes a third dichroic mirror 1307, which is disposed in the optical path between the first light source 11a, the second light source 11b, and the dichroic mirror 1301. The third dichroic mirror 1307 is configured to reflect the first detection light and transmit the second detection light. By setting the third dichroic mirror 1307, the laser direction can be adjusted so that the two detection lights emitted by the first light source 11a and the second light source 11b are emitted in the same direction, thereby allowing both the first and second detection lights to enter the objective lens 40.

[0151] When the first optical system also includes a third dichroic mirror 1307, the first optical system also includes a second reflecting mirror 1304. The second reflecting mirror 1304 is disposed in the optical path between the second light source 11b and the third dichroic mirror 1307. The second reflecting mirror 1304 is configured to reflect the second detection light to the third dichroic mirror 1307. By setting the second reflecting mirror 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. This is beneficial to keep the first detection light and the second detection light entering the objective lens 40 in different directions, and thus facilitates a more flexible arrangement of the first light source 11a and the second light source 11b.

[0152] Based on the above structure, the light source in this embodiment can be arranged as follows: 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. Thus, 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 devices more compact and helping to reduce the space occupied by the first optical system.

[0153] When the particle pairing device also includes an objective lens 40, optionally, in the direction from the first particle channel to the pairing region, the first detection light (see Figure 13 In the direction from the second particle channel to the pairing area, the second detection light (see S1) is perpendicular to the first particle channel (at this time, the first detection light is perpendicular to the page), and the second detection light (see S1) is perpendicular to the first particle channel (at this time, the first detection light is perpendicular to the page). Figure 13 S2) is perpendicular to the second particle channel (at this time, the second detection light is perpendicular to the page). This helps to ensure that the first particle can be illuminated by the first detection light when it passes through the first particle channel, and ensures that the second particle can be illuminated by the second detection light when it passes through the second particle channel, thereby improving the detection accuracy of the detection module 10 for the first and second particles, and thus improving the reliability of particle pairing.

[0154] When the particle pairing device also 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 channel, nor does the second detection light intersect with the first particle channel. This ensures 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. This helps reduce signal interference and avoids the situation where the first particle is simultaneously affected by both the first and second detection lights, resulting in an interference peak. This further improves the accuracy of the particle pairing device 1 in detecting the first and second particles. More specifically, both the first and second detection lights enter the objective lens 40 in a direction parallel to the optical axis of the objective lens 40.

[0155] When the particle pairing device also 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 does not intersect with the second particle channel, nor does the second detection light intersect with the first particle channel. This ensures 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. This helps reduce signal interference and avoids the situation where the first particle is simultaneously affected by both the first and second detection lights, resulting in an interference peak. This further improves 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 where the first and second detection lights enter the objective lens 40 can be the same or different.

[0156] When the particle pairing device also includes an objective lens 40, in some embodiments, the detection module 10 further includes an optical path adjustment mechanism 13. The optical path adjustment mechanism 13 is disposed 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. This can improve the structural compactness while ensuring that the first detection light and the second detection light irradiate the first particle flow channel and the second particle flow channel, and that 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 reduce the volume of the particle pairing device 1.

[0157] Optionally, the optical path adjustment mechanism 13 includes the aforementioned multi-directional mirror 1301. The multi-directional mirror 1301 is disposed on the side of the objective lens 40 away from the detection window 31. The multi-directional mirror 1301 is used to reflect the laser emitted by the laser emitting mechanism 11 to the objective lens 40 and to transmit 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 improve the reusability of the structure and further improve the structural compactness.

[0158] Optionally, refer to Figure 2 The optical path adjustment mechanism 13 also includes a first mounting bracket 1302 and a second mounting bracket 1303 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, while 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 the adjustment of the directions in which the first and second detection lights enter the objective lens, satisfying the design that the first and second detection lights enter the objective lens 40 at different incident angles.

[0159] Furthermore, the first mounting bracket 1302 and the second mounting bracket 1303 can adjust the laser emission direction of the first light source 11a and the second light source 11b through structural adjustment to change the emission direction of the first detection light and the second detection light from the laser generator. Alternatively, they can be configured with optical elements inside to change the emission direction of the first detection light and the second detection light through the first mounting bracket 1302 and the second mounting bracket 1303. The specific configuration can be set according to actual needs and is not limited here.

[0160] Optionally, the optical path adjustment mechanism 13 further includes the aforementioned second reflector 1304, which is disposed in the first detection light emission direction of the first light source 11a and is used to adjust the optical path direction of the first detection light.

[0161] In this way, 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 beneficial to make the first detection light and the second detection light enter the objective lens 40 in different directions.

[0162] Optionally, refer to Figure 2 , Figure 11 The optical path adjustment mechanism 13 also includes a first cylindrical mirror 1305 and a second cylindrical mirror 1306. The first cylindrical mirror 1305 is disposed between the second reflecting mirror 1304 and the multi-color mirror 1301, and the second cylindrical mirror 1306 is disposed between the second mounting bracket 1303 and the multi-color mirror 1301. The first and second detection lights are converted into strip lasers by the first cylindrical mirror 1305, the second cylindrical mirror 1306, and the objective lens 40 to adjust the image height of the first and second detection lights, thereby adjusting the illumination area of ​​the first and second detection lights to facilitate the detection and identification of the first and second particles in the designated area.

[0163] Optionally, the optical path adjustment mechanism 13 also includes the aforementioned third dichroic mirror 1307. The third dichroic mirror 1307 is disposed 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.

[0164] Optionally, refer to Figure 2The optical path adjustment mechanism 13 also includes a third reflecting mirror 1308 and a fourth reflecting mirror 1309. The third reflecting mirror 1308 and the fourth reflecting mirror 1309 are sequentially and alternately arranged between the third dichroic mirror 1307 and the multi-directional mirror 1301. The third reflecting mirror 1308 and the fourth reflecting mirror 1309 reverse the optical path direction of the first and second detection lights, facilitating the adjustment of the direction of the laser entering the objective lens 40 and simultaneously reducing 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 reflecting mirror 1308 are all located at the bottom layer. The fourth reflecting mirror 1309 and the multi-directional mirror 1301 are located in the middle layer, that is, above the first light source 11a, the second light source 11b, the third dichroic mirror 1307, and the third reflecting mirror 1308. Specifically, the fourth reflecting mirror 1309 is located directly above the third reflecting mirror 1308. The objective lens 40 is located in the upper layer, specifically above the multi-directional mirror 1301. Directly above the dichroic mirror 1301, the detection light passes through the third dichroic mirror 1307 and then enters the third reflecting mirror 1308. After being reflected by the third reflecting mirror 1308, it enters the fourth reflecting mirror 1309 and is then reflected by the fourth reflecting mirror 1309 before entering the multi-color mirror 1301 on the same layer. After being reflected by the multi-color mirror 1301, it enters the objective lens 40. In this way, the horizontal and vertical space can be fully utilized to arrange various optical devices, thus improving space utilization.

[0165] Optionally, the optical path adjustment mechanism 13 also includes a multicolor narrowband filter, which is disposed between the multicolor mirror 1301 and the fourth reflecting mirror 1309. The multicolor narrowband filter filters out some wavelengths of light beams to ensure the purity of the light beam entering the multicolor mirror 1301, thereby improving the purity of the light beam irradiating the first particle channel and the second particle channel, which is beneficial for the detection of the first particle and the second particle.

[0166] Optionally, refer to Figure 4 , Figure 12 The first detection device 12a has a first filter 1312 at its light inlet and the second detection device 12b has a second filter 1313 at its light inlet, which can limit the passage of light beams of specific wavelengths and filter out other stray light, thereby improving the purity of photoelectric signals.

[0167] For example, the first filter 1312 is a 525nm narrowband filter and the second filter 1313 is a 655nm long-pass filter, so that light beams with wavelengths near 525nm enter the first detection device 12a and light beams with wavelengths longer than 655nm enter the second detection device 12b.

[0168] Understandably, in other embodiments, the second dichroic mirror 1311 and the first dichroic mirror 15 may also be short-pass dichroic mirrors, so that light beams with wavelengths lower than the cutoff wavelength can pass through, while light beams with wavelengths higher than the cutoff wavelength are reflected.

[0169] In some embodiments, reference is made to Figure 1 The particle pairing device 1 also includes a bright field lamp 50, which illuminates the detection window 31. The bright field lamp 50 is positioned on the side of the detection window 31 away from the objective lens 40, and the bright field lamp 50 and the objective lens 40 are located on opposite sides of the detection window 31, which improves structural rationality and helps to reduce the size of the particle pairing device 1. The light emitted by the bright field lamp 50 passes sequentially through the detection window 31, the objective lens 40, the multi-color mirror 1301, the first reflecting mirror 1310, and the first dichroic mirror 15 into the image generation component 14, thereby illuminating the surface of the microfluidic chip 2. This facilitates the introduction of bright field 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, making it easier for the operator to observe the microfluidic chip 2.

[0170] Optionally, the wavelength of the light emitted by the bright field lamp 50 is different from the wavelength of the first excitation light and the second excitation light. This helps to distinguish the light emitted by the bright field lamp 50 from the first excitation light and the second excitation light, and to drive light of different wavelengths into different optical elements to act. This avoids the situation where the light emitted by the bright field lamp 50 accidentally enters the first detection device 12a or the second detection device 12b, which would lead to the detection error of the first particle and the second particle.

[0171] Furthermore, as mentioned above, the second dichroic mirror 1311 is a 550nm long-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 conditions for being reflected by the second dichroic mirror 1311, so as to ensure that it can enter the image generation component 14.

[0172] Optionally, a narrowband filter is provided at the light outlet of the bright field lamp 50, which can help filter the light emitted by the bright field lamp 50, so that the wavelength of the light emitted through the bright field lamp 50 and the narrowband filter is the target wavelength, thereby reducing the influence of the light from the bright field lamp 50 on the particle detection results.

[0173] To facilitate reading and understanding, the following will combine... Figure 12 A simple example illustrating the motion path of the beam in particle pairing device 1:

[0174] The wavelength of the first detection light is 488nm, and the wavelength of the second detection light is 640nm. After being emitted from the first light source 11a, the first detection light sequentially 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 polarizing beam splitter cube 61, the multicolor narrowband filter, the multicolor mirror 1301, the quarter-wave plate 62, the objective lens 40, and the detection window 31 before entering the microfluidic chip 2. After being emitted from the second light source 11b, the second detection light sequentially 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 polarizing beam splitter cube 61, the multicolor narrowband filter, the multicolor mirror 1301, the quarter-wave plate 62, the objective lens 40, and the detection window 31 before entering the microfluidic chip 2. In the microfluidic chip 2, a first detection light excites a first particle to emit a first excitation light, and a second detection light excites a second particle to emit a second excitation light. The bright field light emitted by the bright field lamp 50, along with the first and second excitation lights, sequentially passes through the detection window 31, objective lens 40, quarter-wave plate 62, dichroic mirror 1301, and first reflector 1310, 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 generation component 14. The first and second excitation lights pass through the first dichroic mirror 15. Subsequently, 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. The second excitation light sequentially passes through the second dichroic mirror 1311 and the second filter 1313 to enter the second detection device 12b.

[0175] For ease of understanding, please refer to the following: Figures 14 to 17 and combined Figures 1 to 3 The particle pairing device 1 involved in this application will be described.

[0176] Reference Figures 15 to 17 and combined 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 drives 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 on the microfluidic chip 2, and also facilitates adjusting the focal length of the objective lens 40, thereby improving the imaging quality of the image generation component 14. Specifically, the three-axis motion mechanism 70 can drive the support platform 30 to move along a first horizontal direction (e.g., the front-to-back direction), a second horizontal direction (e.g., the left-to-right direction), and a vertical direction. For example, 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 lead screw, and a lead screw nut.

[0177] Optionally, refer to Figure 1 and Figure 17 The support platform 30 also includes a focusing knob 32 for adjusting the focal length of the objective lens 40, thereby improving the imaging quality of the image generation component 14. For example, the focusing knob 32 extends from the upper side of the support platform 30 for easy user operation.

[0178] Optionally, refer to Figure 1 The support platform 30 is provided with a liquid storage tank 33, which stores a first particle solution, a second particle solution, a buffer solution and an oil. The liquid storage tank 33 is used to introduce the first particle solution into the first particle channel, introduce the second particle solution into the second particle channel, and introduce the buffer solution and oil into the pairing area.

[0179] In this way, by setting up a liquid storage tank 33 to store the solution required for the pairing process of the first particle and the second particle, it is beneficial to shorten the liquid supply path to the microfluidic chip 2, thereby improving 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 tank, thereby improving the portability of the particle pairing device 1.

[0180] Optionally, refer to Figure 2 The support platform 30 is provided with multiple sample collection slots 34 and multiple sample collection tubes 35. The multiple sample collection slots 34 correspond one-to-one with the multiple 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.

[0181] This facilitates the storage of droplets generated after the first and second particles pair up, reducing the time required for operators to handle droplets and thus improving the efficiency of single-cell sequencing.

[0182] It should be noted that the aforementioned embodiment uses the support platform 30 as the installation base to install components such as the microfluidic chip 2, focusing knob 32, liquid storage tank 33, collection tank 34 and multiple sample collection tubes 35, which can achieve integrated installation and facilitate user operation.

[0183] In some embodiments, reference is made to Figure 1 The particle pairing device 1 also includes a control panel 80, which is electrically connected to the first particle flow channel control valve and the second particle flow channel control valve 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 enables manual control of the first and second particle flow channel control valves, allowing for 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, controlling the start and stop of the valve control module 20 via the control panel 80 can also be used to detect whether the valve control module 20 is functioning properly, thus improving the reliability of single cell-to-microsphere pairing.

[0184] Optionally, the valve control module 20 also includes a control circuit board. 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 electrically connected to the control circuit board. The control panel 80 includes a display screen and control buttons. The display screen and control buttons are electrically connected to the control circuit board. The control circuit board is located on the side of the control panel 80 opposite to 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. There are multiple control buttons, which are 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 opening and closing of the first particle flow channel control valve and the pressure of the second particle flow channel control valve, thereby improving the flexibility of the valve control module 20 and enabling the particle pairing device 1 to freely adjust parameters according to user needs, thereby improving 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 encapsulation program. The specific settings can be configured according to actual needs, and there are no restrictions here.

[0185] Optionally, the control panel 80 also includes an autofocus button, which is used to adjust the focal length of the objective lens 40 by electrical control to achieve autofocus, thereby improving the imaging sharpness of the image generation component 14.

[0186] In some embodiments, reference is made to Figure 1 The particle pairing device 1 also includes a housing and a base 90 disposed at the bottom of the housing. The three-axis motion mechanism 70, the detection module 10 and the valve control module 20 are all disposed on the base 90 and located inside the housing. The control panel 80 is mounted on the housing and is at least partially located outside the housing. Multiple structures of the detection module 10 are arranged around the three-axis motion mechanism 70, so as to make reasonable use of space, which is conducive to ensuring the volume of the particle pairing device 1 while making the detection module 10 have a longer optical path, so as to facilitate the projection imaging of laser and beam.

[0187] In some embodiments, reference is made 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 end of the column 92. For example, the connecting seat 91 is located laterally to the top end of the column 92 and is connected to the top end 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 has 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 a multi-color mirror 1301; for example, the multi-color mirror 1301 is connected to the lower side of the mounting seat 91. In some embodiments, the column 92 is also used to mount a bright field lamp 50. Exemplarily, the bright field lamp 50 includes a lamp post 51 and a bright field light source 52. The lamp post 51 is connected to the top of the column 92, and the bright field light source 52 is connected to the top of the lamp post 51 and located to the side of the lamp post 52, so that the axis of the bright field light source 51 is coaxial with the objective lens 40. In some embodiments, the connector 91 is also used to connect 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.

[0188] In some embodiments, reference is made to Figure 15 , Figure 16The laser emitting mechanism 11, the second reflector 1304, the first cylindrical mirror 1305, the second cylindrical mirror 1306, the third dichroic mirror 1307, the third reflector 1308, the fourth reflector 1309, and the multi-color mirror 1301 are arranged around the three-axis motion mechanism 70. The first light source 11a, the second light source 11b, the second reflector 1304, the first cylindrical mirror 1305, the second cylindrical mirror 1306, the third dichroic mirror 1307, the third reflector 1308, the first detection device 12a, the second detection device 12b, the first reflector 1310, the second dichroic mirror 1311, and the first dichroic mirror 15 are located at the same height. All or part of them are connected to the base 90 through vertically arranged connecting columns to serve as the bottom component of the optical path structure of the particle pairing device 1. The fourth reflecting mirror 1309, the polarizing beam splitter cube 61, and the multi-directional mirror 1301 are located at the same height, with the fourth reflecting mirror 1309 connected above the third reflecting mirror 1308 and the multi-directional mirror 1301 connected above the first reflecting mirror 1310, serving as the middle component of the optical path structure of the particle pairing device 1. The objective lens 40 is connected above the multi-directional mirror 1301, serving as the upper component of the optical path structure of the particle pairing device 1.

[0189] Based on the first embodiment, in some embodiments the detection module 10 includes a second optical system, and the particle pairing device 1 further includes an objective lens 40. The second optical system is configured to receive imaging light from the microfluidic chip 2 and passing through the objective lens 40 to form an image of the microfluidic chip 2. 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 foregoing embodiments. This embodiment mainly protects the scheme of using the image magnification function of the objective lens 40.

[0190] Secondly, this application also provides a particle pairing system (not shown), including a microfluidic chip 2 and a particle pairing device 1 as described in the first aspect above. The microfluidic chip 2 is provided with a pairing region and a first particle flow channel and a second particle flow channel that connect the pairing region, and the microfluidic chip 2 is installed on the particle pairing device 1.

[0191] It is understood that since the particle pairing system includes the particle pairing device 1 described in the first aspect above, the particle pairing system has the beneficial effects of the particle pairing device 1 described in the first aspect above, which will not be repeated here.

[0192] To facilitate reading and understanding, the first particle is represented as a cell and the second particle as a microsphere. The following is a simplified example illustrating the workflow of the particle pairing system:

[0193] Before the particle pairing system starts working, the microfluidic chip 2 is first installed on the support platform 30 using the clamp 36. The air in the flow channels of the microfluidic chip 2 (including the first particle flow channel, the second particle flow channel, and the pairing area, etc.) is emptied. The imaging clarity of the image generation component 14 is adjusted by the focusing knob 32. The control valves of the valve control module 20 are controlled to work sequentially by the control buttons on the control panel 80 to detect whether the valve control module 20 can work normally.

[0194] After the preliminary work is completed, the first light source 11a and the second light source 11b are first activated. Then, the cell solution in the storage tank 33 is introduced into the first particle flow channel and the microsphere solution is introduced into the second particle flow channel using the valve control module 20. 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 and emits a first excitation light, which 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 stopped from entering the first particle flow channel, and the cell solution that has already entered the first particle flow channel is blocked. Cells in the first particle channel, under inertia, flow through the channel to the pairing area and then stop moving. When a microsphere passes through the second particle channel and is irradiated by the second detection light, the microsphere is excited by the second detection light and emits a second excitation light, which is received by the second detection device 12b. The second detection device 12b controls the second particle channel control valve to block the second particle channel. At this time, the microsphere solution is stopped, and entry into the second particle channel is halted. Microspheres that have already entered the second particle channel, under inertia, flow through the channel to the pairing area and then stop moving. Subsequently, the buffer solution in the reservoir 33 is introduced into the pairing area through the valve control module 20, so that the buffer solution brings the cells and microspheres closer or into contact. Then, the oil in the reservoir 33 is introduced into the pairing area through the valve control module 20, so that the oil co-encapsulates the cells and microspheres, forming droplets of single cells paired with single microspheres. These droplets are discharged through the sample outlet of the microfluidic chip 2 to the sample collection tube 35.

[0195] 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 for the next round of cell and microsphere recognition and pairing.

[0196] Thirdly, this application also provides an optical module for identifying a first particle and a second particle. The module includes an objective lens 40, a first optical system, and a second optical system. The first optical system generates a first detection light passing through the objective lens 40 and a second detection light passing through the objective lens 40. The first particle can be illuminated by the first detection light to emit a first excitation light, and the second particle can be illuminated by the second detection light to emit a second excitation light. The second optical system receives imaging light from the microfluidic chip 2 and passing through the objective lens 40 to form images of the first and / or second particles. This allows users to directly check whether the particles have reached the corresponding positions within the flow channel.

[0197] 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 the small structural size of the microfluidic chip 2, if the spot area is large, it may cover some unsuitable positions, such as covering another particle channel, thus leading to detection errors. That is, the detection module 10 has high requirements for the precision of the spot when detecting the first and second particle channels. On the premise of being able to cover the detection position of the corresponding particle channel, reducing the area of ​​the spot is beneficial to reducing interference. Based on this, the adjustment of the spot size mentioned 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 illuminate the first and second particle channels, and make the size of the detection light of the detection module 10 adapt to the size of the first and second particle channels, thereby facilitating the detection module 10 to detect the first and second particle channels.

[0198] Furthermore, the objective lens 40 in this embodiment can also be applied to the imaging system to assist in magnifying the image. As mentioned earlier, the microfluidic chip 2 has a small structural size, and the first particle channel, the second particle channel, the first particle, and the second particle are difficult to see clearly with the naked eye. Therefore, the imaging light in this embodiment will also pass through the objective lens 40 to magnify the image, so that the user no longer needs to use other tools to see the first particle channel, the second particle channel, the first particle, and the second particle.

[0199] Based on the third aspect of the embodiments, in some embodiments of this application, the objective lens 40 can also be used for shaping the light spot. Specifically, the cross-section of the light beam emitted by the light source is usually rectangular or circular, with a large coverage area, and its shape is not suitable for the 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 illuminating the microfluidic chip 2 is a strip-shaped light spot (e.g., Figure 13As shown), at this time, the objective lens 40 is configured to at least adjust the corresponding size of the strip-shaped light spot. For example, the objective lens 40 is configured to adjust the width of the strip-shaped light spot so that the light spot can be concentrated at the detection position. Or, for example, the objective lens 40 is configured to adjust the length of the strip-shaped light spot so as to adapt to particle channels of different widths.

[0200] Based on the third aspect of the embodiments, in some embodiments of this application, reference is made 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 into the first particle channel through the objective lens 40 and the detection window 31 in sequence, and can also emit a second detection light into the second particle channel. The first detection light is used to cause the first particle in the first particle channel to emit a first excitation light, and the second detection light is used to cause the second particle in the second particle channel to emit a second excitation light. The detection component 12 is used to receive the first excitation light and control the first particle channel control valve to block the first particle channel. The detection component 12 is also used to receive the second excitation light and control the second particle channel control valve to block the second particle channel.

[0201] In this way, by setting up the laser emitting mechanism 11, the first particle and the second particle emit the first excitation light and the second excitation light respectively under the action of the laser, so that the detection component 12 controls the first particle flow channel control valve and the second particle flow channel control valve respectively under the control of the corresponding 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.

[0202] Understandably, in other embodiments, the detection module 10 may also use electrode detection or visual image recognition to detect the first particle and the second particle.

[0203] The second optical system includes an image generation component 14, which receives imaging light and forms an image. This facilitates the operator's observation of the microfluidic chip 2's operating status, improving operational convenience and enabling visual detection and manual identification of the first and second particles. This allows the particle pairing device 1 to employ different detection methods for the first and second particles, enriching its functionality. Furthermore, visual detection or manual identification can interlock the functions of the detection module 10 and the valve control module 20, improving the reliability of the particle pairing device 1 in matching the first and second particles and thus enhancing the quality of particle pairing. The image generation component 14 may include a camera and a display screen.

[0204] In this embodiment, the first optical system and the second optical system include a first shared optical path. The first shared optical path is roughly marked in the figure by the area enclosed by a long dashed line. Specifically, the first shared optical path refers to the conduit portion shared by the excitation light and the imaging light. (Refer to...) Figure 2 , Figure 3 It includes a multi-color mirror 1301, which is an optical element capable of allowing or reflecting three or more different wavelengths or wavelength ranges of light. In this embodiment, the multi-color mirror 1301 is configured to reflect the first detection light and the second detection light to the objective lens 40, and transmit the first excitation light, the second excitation light, and the imaging light. Thus, by using a single optical element to achieve the reflection and transmission of different lights, the reusability of the structure is improved, further enhancing the structural compactness. It should be noted that the multi-color mirror 1301 can employ known technologies.

[0205] Based on the third aspect of the embodiments, in some embodiments of this application, the first optical system further includes a detection component 12, which can be understood with reference to the foregoing embodiments.

[0206] Reference Figure 1 , Figure 8 The first shared optical path also includes a first dichroic mirror 15, which is disposed in the optical path between the multi-directional mirror 1301 and the detection component 12. The first dichroic mirror 15 is configured to reflect the imaging light emitted from the multi-directional mirror 1301 to the image generation component 14, and to transmit the first excitation light and the second excitation light emitted from the multi-directional mirror 1301 to the detection component 12. Thus, by setting the first dichroic mirror 15, the first and second excitation lights can pass through the first dichroic mirror 15 to facilitate their subsequent entry into the detection component 12, while other wavelengths of imaging light can be utilized to generate an image through the image generation component 14, facilitating the observation of the working state of the microfluidic chip 2. It should be noted that the first dichroic mirror 15 can employ known technology.

[0207] As an alternative to the aforementioned embodiment, the first dichroic mirror 15 is configured to transmit the imaging light emitted after passing through the dichroic mirror 1301 to the image generation component 14, and reflect the first excitation light and the second excitation light emitted after passing through the dichroic mirror 1301 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 reversed.

[0208] Based on the third aspect of the embodiments, in some embodiments of this 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 and 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 and is used to receive the second excitation light and control the second particle flow channel control valve to block the second particle flow channel.

[0209] In this embodiment, refer to Figure 1 , Figure 7 The first optical system further includes a second dichroic mirror 1311, which is disposed in the optical path between the multi-directional mirror 1301 and the detection component 12. The second dichroic mirror 1311 is configured to reflect the first excitation light emitted after passing through the multi-directional mirror 1301 to the first detection device 12a, and transmit the second excitation light emitted after passing through the multi-directional mirror 1301 to the second detection device 12b.

[0210] 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 sequentially through the objective lens 40, the multi-color mirror 1301 and the second dichroic mirror 1311, and the second excitation light enters the second detection device 12b sequentially through the objective lens 40, the multi-color mirror 1301 and the second dichroic mirror 1311.

[0211] In this way, the second dichroic mirror 1311 makes the first excitation light and the second excitation light have different transmission directions, so that the two 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 signal interference between the two beams, avoiding signal crossing, and thus improving the control accuracy of the first particle flow channel and the second particle flow channel.

[0212] The embodiments of this application have been described in detail above with reference to the accompanying drawings. However, this application is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of this application. Furthermore, unless otherwise specified, the embodiments and features described in the embodiments of this application can be combined with each other.

Claims

1. A particle pairing system, characterized in that, Including microfluidic chips and particle pairing devices: The microfluidic chip includes a first particle channel, a second particle channel, an oil channel, a buffer solution channel, and a paired channel. The paired channel is connected to the oil channel, and the first particle channel, the second particle channel, and the buffer solution channel are all connected to the paired channel. The microfluidic chip further includes a pairing microvalve corresponding to the paired flow channel, a first microvalve corresponding to the first particle flow channel, a second microvalve corresponding to the second particle flow channel, and a buffer microvalve corresponding to the buffer flow channel. The pairing microvalve is used to control the opening and closing of the paired flow channel, the first microvalve is used to control the opening and closing of the first particle flow channel, the second microvalve is used to control the opening and closing of the second particle flow channel, and the buffer microvalve is used to control the opening and closing of the buffer flow channel. The particle pairing device includes a detection module and a valve control module. The detection module is used to identify a first particle when the first particle solution flows through the first particle channel, and the detection module is also used to identify a second particle when the second particle solution flows through the second particle channel. The valve control module is electrically connected to the detection module and is used to block the first particle channel when the detection module identifies the first particle, and to block the second particle channel when the detection module identifies the second particle. The detection module includes a first optical system, and the particle pairing device further includes an objective lens. The first optical system is used to generate a first detection light that passes through the objective lens and illuminates the first particle channel, and to generate a second detection light that passes through the objective lens and illuminates the second particle channel. The first particle can be illuminated by the first detection light to emit a first excitation light, and the second particle can be illuminated by the second detection light to emit a second excitation light. The first detection light and the second detection light enter the objective lens at different incident angles, so that the spot of the first detection light illuminates the first particle flow channel and the spot of the second detection light illuminates the second particle flow channel. The spot is a strip-shaped spot. The objective lens is configured to at least adjust the width of the strip-shaped spots of the first detection light and the second detection light, so that the strip-shaped spot of the first detection light is concentrated at the detection position of the first particle flow channel and the strip-shaped spot of the second detection light is concentrated at the detection position of the second particle flow channel. When a single first particle and a single second particle are present in the paired flow channel, the first microvalve and the second microvalve are in a closed state, and the paired microvalve and the buffer microvalve are in an open state, so that the buffer in the buffer flow channel can deliver the single first particle and the single second particle in the paired flow channel into the oil in the oil flow channel to form droplets.

2. The particle pairing system according to claim 1, characterized in that, The objective lens is configured to at least adjust the length of the stripe-shaped light spots of the first detection light and the second detection light.

3. The particle pairing system according to claim 1, characterized in that, The detection module includes a second optical system 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.

4. The particle pairing system according to claim 3, characterized in that, The objective lens is configured such that the directions in which the first detection light and the second detection light pass through the objective lens are opposite to the directions in which the imaging light passes through the objective lens.

5. The particle pairing system according to claim 3, 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 includes an image generation component, which 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, which includes a multi-directional color mirror. The multi-directional color 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.

6. The particle pairing system according to claim 5, characterized in that, The first optical system further includes a detection component, which is used to receive the first excitation light and the second excitation light, and to identify the first particle based on the first excitation light and 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 multi-directional mirror and the detection component. The first dichroic mirror is configured to reflect the imaging light emitted from the multi-directional mirror to the image generation component, and to transmit the first excitation light and the second excitation light emitted from the multi-directional mirror to the detection component. Alternatively, the first dichroic mirror is configured to transmit the imaging light emitted after passing through the dichroic mirror to the image generation component, and to reflect the first excitation light and the second excitation light emitted after passing through the dichroic mirror to the detection component.

7. The particle pairing system according to claim 5, characterized in that, The first optical system further includes a detection component, which includes a first detection device and a second detection device. The first detection device is used to receive the first excitation light and identify the first particle based on the first excitation light. The second detection device is used to receive the second excitation light and identify the second particle based on the second excitation light. The first optical system further includes a second dichroic mirror, which is disposed in the optical path between the multi-directional mirror and the detection component. The second dichroic mirror is configured to reflect the first excitation light emitted after passing through the multi-directional mirror to the first detection device, and to transmit the second excitation light emitted after passing through the multi-directional mirror to the second detection device.

8. The particle pairing system according to claim 5, characterized in that, The first shared optical path further includes a first reflector, which is disposed on the optical path between the multi-directional color mirror and the image generation component. The first reflector is configured to reflect the first excitation light, the second excitation light and the imaging light emitted after passing through the multi-directional color mirror.

9. The particle pairing system according to claim 8, characterized in that, The objective lens, the multi-color mirror, and the first reflecting mirror are arranged vertically. The first reflecting mirror is configured such that the first excitation light, the second excitation light, and the imaging light emitted after passing through the multi-color mirror are directed vertically toward the first reflecting mirror and emitted horizontally after being reflected by the first reflecting mirror.

10. The particle pairing system according to claim 3, 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 disposed in the optical path between the objective lens and the image generating component.

11. The particle pairing system according to claim 10, characterized in that, The first optical system includes a first light source and a second light source, wherein 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 multi-directional color mirror disposed in the optical path between the objective lens and the image generating component. The multi-directional color 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. The first optical system further includes a polarization beam splitter cube, which is disposed in the optical path between the first light source, the second light source and the multi-directional color mirror.

12. The particle pairing system according to claim 1, characterized in that, The first optical system includes a first light source, a second light source, and a multi-directional 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 multi-directional mirror is disposed in the optical path between the first light source, the second light source, and the objective lens. The multi-directional mirror is configured to reflect the first detection light and the second detection light to the objective lens, and transmit the first excitation light emitted by the first particle after being irradiated by the first detection light, and the second excitation light emitted by the second particle after being irradiated by the second detection light.

13. The particle pairing system according to claim 12, characterized in that, The first optical system further includes a third dichroic mirror, which is disposed in the optical path between the first light source, the second light source and the dichroic mirror. The third dichroic mirror is configured to reflect the first detection light and transmit the second detection light.

14. The particle pairing system according to claim 13, characterized in that, The first optical system further includes a second mirror, which is disposed in the optical path between the second light source and the third dichroic mirror, and is configured to reflect the second detection light to 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.

15. The particle pairing system according to claim 1, characterized in that, The wavelengths of the first detection light and the second detection light are different.

16. The particle pairing system according to claim 1, characterized in that, The microfluidic chip has a pairing region, and both the first particle channel and the second particle channel are connected to the pairing region. In the direction from the first particle channel to the pairing region, the first detection light is perpendicular to the first particle channel, and in the direction from the second particle channel to the pairing region, the second detection light is perpendicular to the second particle channel.

17. The particle pairing system according to any one of claims 1 to 16, characterized in that, The particle pairing device further includes a support platform for mounting the microfluidic chip, and the objective lens is disposed on the other side of the support platform, which is different from the microfluidic chip.

18. The particle pairing system according to claim 17, characterized in that, The microfluidic chip has a pairing area, and the first particle channel and the second particle channel are both connected to the pairing area. The support platform is provided with a detection window corresponding to the microfluidic chip. The detection window is connected to the 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 channel, and the second particle 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.

19. The particle pairing system according to claim 17, characterized in that, The particle pairing device further includes a three-axis motion mechanism, on which the support platform is mounted. The three-axis motion mechanism is used to drive the support platform to move in three mutually perpendicular directions.

20. The particle pairing system according to claim 1, characterized in that, The detection module includes a second optical system 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.

21. The particle pairing system according to any one of claims 1 to 16, characterized in that, The microfluidic chip has a pairing region, and the first particle channel and the second particle channel are both connected to the pairing region. The particle pairing device is provided with a liquid storage tank, which stores a first particle solution, a second particle solution, a buffer solution, and an oil. The liquid storage tank is used to introduce the first particle solution into the first particle channel, introduce the second particle solution into the second particle channel, and introduce the buffer solution and the oil into the pairing region.

22. The particle pairing system according to any one of claims 1 to 16, characterized in that, The particle pairing device is provided with multiple sample collection slots and multiple sample collection tubes. Each sample collection slot corresponds to one of the multiple 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.

23. The particle pairing system according to any one of claims 1 to 16, characterized in that, The particle pairing device also 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.

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