Microfluidic system and pairing method for target pairing

Abstract

The application discloses a micro-fluidic system and a pairing method for target pairing. The micro-fluidic system comprises a first flow channel, a second flow channel, an oil flow channel, a buffer flow channel, a pairing flow channel, a pairing micro-valve, a first micro-valve, a second micro-valve and a buffer micro-valve. The pairing flow channel is communicated to the oil flow channel. The first flow channel, the second flow channel and the buffer flow channel are all communicated to the pairing flow channel. When there is a single first target and a single second target in the pairing flow channel, the first micro-valve and the second micro-valve are in a closed state, and the pairing micro-valve and the buffer micro-valve are in an open state. The buffer in the buffer flow channel sends the single first target and the single second target in the pairing flow channel into the oil in the oil flow channel to form a droplet. The application can completely avoid other first targets and / or other second targets from entering the droplet, which is beneficial to improving the success rate of encapsulation and reducing cell loss caused by multiple encapsulation.

Description

Technical Field

[0001] This invention relates to the field of single-cell sequencing technology, and in particular to a microfluidic system and pairing method for target pairing. Background Technology

[0002] Single-cell RNA sequencing (scRNA-seq) technology reveals the heterogeneity of cells within tissues. Compared to traditional gene sequencing technologies, single-cell sequencing offers advantages such as high accuracy and high resolution, providing information at the single-cell level and revealing the heterogeneity and complexity of expression within tissues. It has wide applications in fields such as cancer, reproduction, immunity, and development. Figure 1 A pairing system for pairing a single first target object with a single second target object is shown. The system includes a first flow channel 1, a second flow channel 2, an oil flow channel 3, a waste liquid flow channel 4, a sample flow channel 5, and multiple microvalves corresponding to each flow channel. The microvalves control the opening and closing of the respective flow channels. The first flow channel 1 has a first stop region 11, and the second flow channel 2 has a second stop region 21. In use, a first liquid containing the first target object is injected into the first flow channel 1, and a second liquid containing the second target object is injected into the second flow channel 2. If no single first target object appears in the first stop region 11, the first liquid is discharged through the waste liquid flow channel 4. Similarly, if no single second target object appears in the second stop region 21, the second liquid is also discharged through the waste liquid flow channel 4. If a single first target object appears in the first stop area 11, the flow of the first liquid in the first flow channel 1 is cut off by the corresponding micro-valve, causing the single first target object to stop in the first stop area 11. Similarly, if a single second target object appears in the second stop area 21, the flow of the second liquid in the second flow channel 2 is cut off by the corresponding micro-valve, causing the single second target object to stop in the second stop area 21. Once both the single first and single second target objects have stopped at their respective stop positions, the waste liquid flow channel 4 is cut off, and the oil flow channel 3 and sample flow channel 5 are opened. Then, the first flow channel 1 and the second flow channel 2 are opened, allowing the single first and single second target objects to be fed into the oil through the first and second liquids respectively, forming droplets. The droplets then flow out through the sample flow channel 5. The above scheme has the following problems:

[0003] 1. The power sources for propelling a single first target object and a single second target object into the oil are the first liquid and the second liquid, respectively. Therefore, the following problems may occur: 1. When the first liquid propels a single first target object, subsequent first target objects will also be encapsulated in the droplet; 2. When the second liquid propels a single second target object, subsequent second target objects will also be encapsulated in the droplet; 3. When the first liquid propels a single first target object, and the second liquid propels a single second target object, subsequent first and subsequent second target objects will be encapsulated in the droplet together, resulting in the formed droplet containing multiple first target objects and / or multiple second target objects, causing multiple encapsulation, which will affect subsequent detection.

[0004] 2. Figure 1 The proposed solution generates a large number of empty droplets, which need to be separated from normal droplets to avoid mixing. Therefore, the processing speed is slow and the efficiency is low. Summary of the Invention

[0005] This invention aims to address at least one of the technical problems existing in the prior art. To this end, this invention proposes a microfluidic system for target pairing.

[0006] The present invention also proposes a pairing method.

[0007] According to a first embodiment of the present invention, a microfluidic system for target pairing includes a first channel, a second channel, an oil channel, a buffer channel, and a pairing channel, wherein the pairing channel is connected to the oil channel, and the first channel, the second channel, and the buffer channel are all connected to the pairing channel;

[0008] The microfluidic system further includes a pairing microvalve corresponding to the paired flow channel, a first microvalve corresponding to the first flow channel, a second microvalve corresponding to the second 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 flow channel, the second microvalve is used to control the opening and closing of the second flow channel, and the buffer microvalve is used to control the opening and closing of the buffer flow channel.

[0009] When a single first target and a single second target exist within 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 delivers the single first target and the single second target in the paired flow channel into the oil in the oil flow channel to form droplets.

[0010] The microfluidic system for target pairing according to embodiments of the present invention has at least the following beneficial effects:

[0011] This embodiment features a separate buffer channel. The buffer channel propels a single first target A and a single second target B into the oil, eliminating the need for the first liquid and the second liquid to propel them. Therefore, the first and second channels can remain in a closed state during the encapsulation process, completely preventing other first targets and / or other second targets from entering the droplet. This improves the success rate of encapsulation and reduces cell loss caused by multiple encapsulations.

[0012] In other embodiments of the present invention, the microfluidic system further includes a waste liquid channel and a waste liquid microvalve corresponding to the waste liquid channel, the waste liquid channel being connected to the paired channel, and the waste liquid microvalve being used to control the opening and closing of the waste liquid channel;

[0013] When the paired microvalve is in the closed state and the first microvalve and the waste liquid microvalve are in the open state, the liquid in the first flow channel can flow out through the paired flow channel and the waste liquid flow channel;

[0014] And / or, when the paired microvalve is in the closed state and the second microvalve and the waste liquid microvalve are in the open state, the liquid in the second flow channel can flow out through the paired flow channel and the waste liquid flow channel.

[0015] In other embodiments of the present invention, the first flow channel, the second flow channel, the buffer solution flow channel, and the waste liquid flow channel are all located on the side of the paired microvalve away from the oil flow channel.

[0016] In other embodiments of the present invention, the first flow channel and the second flow channel are located on the same side of the paired flow channels, and the waste liquid flow channel is located on the other side of the paired flow channels.

[0017] In other embodiments of the present invention, the microfluidic system further includes at least one of the following:

[0018] At least a first portion of the first flow channel that communicates with the paired flow channel is intersected with the paired flow channel;

[0019] The second portion of the second flow channel, which is at least connected to the paired flow channel, is intersecting with the paired flow channel;

[0020] At least a third portion of the waste liquid flow channel that is connected to the paired flow channel is intersected with the paired flow channel.

[0021] In other embodiments of the invention, a fourth portion of the buffer channel that is at least in communication with the paired channel is coaxially disposed with the paired channel.

[0022] In other embodiments of the invention, the waste liquid channel is located between the first channel and the second channel along the extending direction of the paired channels.

[0023] In other embodiments of the present invention, the microfluidic system further includes a detection module configured to identify a single first target object within the first flow channel, wherein when the detection module identifies a single first target object, both the first microvalve and the paired microvalve are in a closed state to keep the single first target object within the paired flow channel;

[0024] And / or, the microfluidic system further includes a detection module configured to identify a single second target object within the second flow channel, wherein, when the detection module identifies a single second target object, both the second microvalve and the paired microvalve are closed to keep the single second target object within the paired flow channel.

[0025] In other embodiments of the present invention, the microfluidic system further includes a waste liquid channel and a waste liquid microvalve corresponding to the waste liquid channel, the waste liquid channel being connected to the paired channel, and the waste liquid microvalve being used to control the opening and closing of the waste liquid channel;

[0026] When the detection module fails to identify a single first target object, the paired microvalve is in a closed state and the first microvalve and the waste liquid microvalve are in an open state, so that the liquid in the first flow channel can flow out through the paired flow channel and the waste liquid flow channel;

[0027] And / or, when the detection module does not identify a single second target, the paired microvalve is in a closed state and the second microvalve and the waste liquid microvalve are in an open state, so that the liquid in the second flow channel can flow out through the paired flow channel and the waste liquid flow channel.

[0028] In other embodiments of the present invention, the detection module includes a camera and a controller, wherein the controller is configured to control the camera to capture images;

[0029] The image includes at least a first image of a first portion of the first flow channel communicating with the paired flow channel, and the controller is further configured to identify the single first target object based on the first image;

[0030] And / or, the image includes at least a second image of a second portion of the second flow channel communicating with the paired flow channel, and the controller is further configured to identify the single second target based on the second image.

[0031] In other embodiments of the invention, the image includes at least a third image of the paired flow channel, and the controller is configured to identify, based on the third image, whether the single first target and / or the single second target exists within the paired flow channel.

[0032] In other embodiments of the present invention, the detection module includes a first light source, a first light detection device, and a controller. The controller is configured to control the first light source to emit a first detection light into the first flow channel. The first detection light can excite a first fluorescence after irradiating the single first target object. When the first light detection device detects the first fluorescence, the detection device identifies the single first target object.

[0033] And / or, the detection module includes a second light source, a second light detection device, and a controller, wherein the controller is configured to control the second light source to emit a second detection light into the second flow channel, the second detection light being able to excite a second fluorescence after irradiating the single second target object, and the controller recognizing the single second target object when the second light detection device detects the second fluorescence.

[0034] In other embodiments of the present invention, the detection module includes a first detection electrode and a controller, the first detection electrode extending into the first flow channel, and the controller being configured to identify the single first target object based on the signal detected by the first detection electrode;

[0035] And / or, the detection module includes a second detection electrode and a controller, the second detection electrode extending into the second flow channel, and the controller being configured to identify the single second target object based on the signal detected by the second detection electrode.

[0036] In other embodiments of the present invention, when the detection module identifies the single first target object, the first microvalve switches from the open state to the closed state after a set delay, so that the single first target object enters the paired flow channel from the first flow channel;

[0037] And / or, when the detection module detects the single second target object, the second microvalve switches from the open state to the closed state after a set delay, so that the single second target object enters the paired flow channel from the second flow channel.

[0038] In other embodiments of the present invention, the microfluidic system further includes an injection channel that is connected to the oil channel. The connection between the paired channel and the oil channel is defined as a first connection point, and the connection between the injection channel and the oil channel is defined as a second connection point. The second connection point is located downstream of the first connection point along the flow direction of the oil in the oil channel.

[0039] The microfluidic system further includes a fusion electrode, which is used to apply an electric field to the second connection portion to cause the liquid in the injection channel to fuse into the droplet flowing through the second connection portion.

[0040] In other embodiments of the present invention, the microfluidic system has a processing area, wherein the first flow channel, the second flow channel, the oil flow channel, the buffer flow channel, the paired flow channel and the injection flow channel are all located within the processing area;

[0041] The fusion electrode includes a first electrode and a second electrode, with the first electrode arranged around the processing area.

[0042] In other embodiments of the present invention, the two ends of the first electrode are arranged side by side with the second electrode, and the second electrode is disposed between the two ends of the first electrode.

[0043] In other embodiments of the present invention, the microfluidic system further includes an oil microvalve disposed corresponding to the oil flow channel, the oil microvalve being used to control the opening and closing of the oil flow channel.

[0044] In other embodiments of the present invention, the microfluidic system further includes a first microvalve control channel, the first microvalve including a diaphragm disposed between the first channel and the first microvalve control channel, the diaphragm being driven to protrude toward the first channel to be in a closed state;

[0045] The inner wall of the first flow channel is an arc-shaped inner wall.

[0046] In other embodiments of the present invention, the microfluidic system includes a microfluidic chip, the microfluidic chip including a base layer, a control layer and a flow channel layer stacked sequentially;

[0047] The flow channel layer includes a first flow channel, a second flow channel, an oil flow channel, a buffer flow channel, and a paired flow channel.

[0048] The control layer includes the paired microvalve, the first microvalve, the second microvalve, and the buffer microvalve.

[0049] In other embodiments of the present invention, the first target is a cell and the second target is a microsphere.

[0050] In other embodiments of the present invention, the microspheres are magnetic microspheres.

[0051] In other embodiments of the present invention, the microspheres are fluorescent magnetic microspheres.

[0052] In other embodiments of the present invention, the first target is a first cell, and the second target is a second cell.

[0053] According to a second embodiment of the present invention, a microfluidic system for target pairing includes a first channel, a second channel, an oil channel, a buffer channel, and a pairing channel. The pairing channel is connected to the oil channel. The first channel and the second channel are both connected to the pairing channel. The buffer channel is connected to both the first channel and the second channel. The first channel has a first portion connected to the pairing channel, and the second channel has a second portion connected to the pairing channel.

[0054] The microfluidic system further includes a first microvalve corresponding to the first flow channel, a second microvalve corresponding to the second flow channel, and a buffer microvalve corresponding to the buffer flow channel. The first microvalve is used to control the opening and closing of the first flow channel, the second microvalve is used to control the opening and closing of the second flow channel, and the buffer microvalve is used to control the opening and closing of the buffer flow channel.

[0055] When a single first target object exists in the first part and a single second target object exists in the second part, the first microvalve and the second microvalve are in a closed state, and the buffer microvalve is in an open state, so that the buffer in the buffer channel delivers the single first target object in the first part and the single second target object in the second part into the oil in the oil channel through the paired channel to form droplets.

[0056] In other embodiments of the present invention, the microfluidic system further includes a waste liquid channel, a droplet channel, a waste liquid microvalve corresponding to the waste liquid channel, and a droplet microvalve corresponding to the droplet channel. The waste liquid channel and the droplet channel are respectively connected to the oil channel. The waste liquid microvalve is used to control the opening and closing of the waste liquid channel, and the droplet microvalve is used to control the opening and closing of the droplet channel.

[0057] When the first micro-valve and the waste liquid micro-valve are in the open state and the droplet micro-valve is in the closed state, the liquid in the first flow channel can flow out through the paired flow channel, the oil flow channel and the waste liquid flow channel;

[0058] And / or, when the second microvalve and the waste liquid microvalve are in the open state and the droplet microvalve is in the closed state, the liquid in the second flow channel can flow out through the paired flow channel, the oil flow channel and the waste liquid flow channel.

[0059] In other embodiments of the present invention, the microfluidic system further includes a waste liquid channel, a droplet channel, a waste liquid microvalve corresponding to the waste liquid channel, and a droplet microvalve corresponding to the droplet channel. The waste liquid channel and the droplet channel are respectively connected to the oil channel. The waste liquid microvalve is used to control the opening and closing of the waste liquid channel, and the droplet microvalve is used to control the opening and closing of the droplet channel.

[0060] When droplets are formed in the oil in the oil flow channel, the waste liquid micro-valve is in a closed state and the droplet micro-valve is in an open state, so that the droplets are discharged through the droplet flow channel.

[0061] In other embodiments of the present invention, the microfluidic system further includes a waste liquid channel, a droplet channel, a waste liquid microvalve corresponding to the waste liquid channel, and a droplet microvalve corresponding to the droplet channel. The waste liquid microvalve is used to control the opening and closing of the waste liquid channel, and the droplet microvalve is used to control the opening and closing of the droplet channel.

[0062] The paired flow channel is connected to the inlet end of the oil flow channel, and the waste liquid flow channel and the droplet flow channel are both connected to the outlet end of the oil flow channel.

[0063] In other embodiments of the present invention, the microfluidic system further includes a detection module configured to identify a single first target object within the first flow channel, wherein when the detection module identifies a single first target object, the first microvalve is closed to keep the single first target object within the first portion;

[0064] And / or, the microfluidic system further includes a detection module configured to identify a single second target object within the second flow channel, wherein, when the detection module identifies a single second target object, the second microvalve is closed to keep the single second target object within the second portion.

[0065] The pairing method according to the third embodiment of the present invention includes the following steps:

[0066] Identify a single first target object from a first liquid carrying a first target object, and stop the first liquid after identifying the single first target object so that the single first target object stops in the target area;

[0067] Identify a single second target object from a second liquid carrying a second target object, and after identifying the single second target object, stop the second liquid so that the single second target object stops in the target area;

[0068] Once both the single first target and the single second target have stopped in the target area, the single first target and the single second target in the target area are introduced into the oil through a third liquid to form droplets.

[0069] In other embodiments of the present invention, the method for identifying a single first target object from a first liquid carrying a first target object includes the following steps: acquiring an image of a detection area, and identifying the single first target object based on image information in the image, wherein the first liquid is capable of entering the target area after passing through the detection area;

[0070] And / or, the method for identifying a single second target object from a second liquid carrying a second target object includes the following steps: acquiring an image of a detection area, identifying the single second target object based on image information within the image, wherein the second liquid is capable of entering the target area after passing through the detection area.

[0071] In other embodiments of the present invention, the method for identifying a single first target object from a first liquid carrying a first target object includes the following steps: detecting a light signal within a detection area, and identifying the single first target object based on the detected light signal, wherein the first liquid is capable of entering the target area after passing through the detection area;

[0072] And / or, the method for identifying a single second target object from a second liquid carrying a second target object includes the following steps: detecting a light signal within a detection area, and identifying the single second target object based on the detected light signal, wherein the second liquid is capable of entering the target area after passing through the detection area.

[0073] In other embodiments of the present invention, the method for identifying a single first target object from a first liquid carrying a first target object includes the following steps: detecting an electrical signal generated by an electrode disposed in a detection area, and identifying the single first target object based on the detected electrical signal, wherein the first liquid is capable of entering the target area after passing through the detection area;

[0074] And / or, the method for identifying a single second target object from a second liquid carrying a second target object includes the following steps: detecting an electrical signal generated by an electrode disposed in a detection area, and identifying the single second target object based on the detected electrical signal, wherein the second liquid is capable of entering the target area after passing through the detection area.

[0075] In other embodiments of the invention, the pairing method further includes the following steps: causing the droplet containing the single first target and the single second target to pass through a fusion region, and applying an electric field to the fusion region to incorporate a fourth liquid into the droplet.

[0076] Additional aspects and advantages of the invention 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 the invention. Attached Figure Description

[0077] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein:

[0078] Figure 1 This is a schematic diagram of a pairing system in related technologies;

[0079] Figure 2 This is a schematic diagram of the microfluidic system in the first embodiment of the present invention;

[0080] Figure 3 for Figure 2 The image shows an enlarged schematic diagram of the paired flow channels;

[0081] Figure 4 This is a schematic diagram of the process by which the microfluidic system achieves pairing between a first target object and a second target object in the first embodiment of the present invention;

[0082] Figure 5 This is a schematic diagram of the microfluidic system combined with the electrode detection module in the first embodiment of the present invention;

[0083] Figure 6 This is a schematic diagram of the microfluidic system combined with the electrofusion module in the first embodiment of the present invention;

[0084] Figure 7 for Figure 6 The image shows an enlarged schematic diagram of the electrofusion process.

[0085] Figure 8 This is a schematic diagram of the microvalve of the microfluidic system in the first embodiment of the present invention in the open and closed states;

[0086] Figure 9 This is an exploded view of the microfluidic chip in the first embodiment of the present invention;

[0087] Figure 10 This is a schematic diagram of the microfluidic system in the second embodiment of the present invention.

[0088] Figure label:

[0089] First flow channel 1, second flow channel 2, oil flow channel 3, waste liquid flow channel 4, sample flow channel 5;

[0090] Microfluidic chip 100, base layer 110, control layer 120, first microvalve 121, diaphragm 1211, second microvalve 122, oil microvalve 123, buffer microvalve 124, waste liquid microvalve 125, paired microvalve 126, flow channel layer 130, first flow channel 131, first part 1311, second flow channel 132, second part 1321, oil flow channel 133, buffer flow channel 134, fourth part 1341, waste liquid flow channel 135, third part 1351, paired flow channel 136, injection flow channel 137, first microvalve control flow channel 138, first injection port 1391, second injection port 1392, third injection port 1393;

[0091] First detection light 210, second detection light 220, first detection electrode 230, second detection electrode 240;

[0092] Fusion electrode 300, first electrode 310, second electrode 320;

[0093] Single primary target A;

[0094] A single second target object B;

[0095] Droplet C. Detailed Implementation

[0096] Embodiments of the present invention 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 the present invention, and should not be construed as limiting the present invention.

[0097] In the description of this invention, it should be understood that the orientation descriptions, such as up, down, front, back, left, right, etc., are based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this invention 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 limiting this invention.

[0098] In the description of this invention, "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.

[0099] In the description of this invention, unless otherwise explicitly defined, terms such as "set up," "install," and "connect" should be interpreted broadly, and those skilled in the art can reasonably determine the specific meaning of the above terms in this invention in conjunction with the specific content of the technical solution.

[0100] In the description of this invention, 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 the invention. 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.

[0101] The first embodiment of the present invention proposes a microfluidic system for target pairing, referring to... Figures 2 to 4 The system includes a first flow channel 131, a second flow channel 132, an oil flow channel 133, a buffer flow channel 134, and a paired flow channel 136. The first flow channel 131 is for the passage of a first liquid containing a first target substance; the second flow channel 132 is for the passage of a second liquid containing a second target substance; the buffer flow channel 134 is for the passage of a buffer solution; the oil flow channel 133 is for the passage of an oil solution that is immiscible with the first liquid, the second liquid, and the buffer solution; and the paired flow channel 136 is for the retention of a single first target substance and a single second target substance. The paired flow channel 136 is connected to the oil flow channel 133, and the first flow channel 131, the second flow channel 132, and the buffer flow channel 134 are all connected to the paired flow channel 136. In some further embodiments, the microfluidic system also includes a waste liquid flow channel 135 for discharging the first liquid and the second liquid. In this embodiment, the first target is a cell and the second target is a microsphere. Those skilled in the art will understand that the first target can be a cell and the second target can also be a cell.

[0102] The microfluidic system in this embodiment also includes multiple microvalves, specifically including a paired microvalves 126 corresponding to the paired flow channel 136, a first microvalves 121 corresponding to the first flow channel 131, a second microvalves 122 corresponding to the second flow channel 132, and a buffer microvalves 124 corresponding to the buffer flow channel 134. The paired microvalves 126 control the opening and closing of the paired flow channel 136, the first microvalves 121 control the opening and closing of the first flow channel 131, the second microvalves 122 control the opening and closing of the second flow channel 132, and the buffer microvalves 124... 124 is used to control the opening and closing of the buffer solution channel 134. Taking the first channel 131 and the first microvalve 121 as an example, the first microvalve 121 controlling the opening and closing of the first channel 131 means that the first microvalve 121 has an open state and a closed state. When the first microvalve 121 is in the open state, the first channel 131 is in the unobstructed state, and the first liquid can flow in the first channel 131. When the first microvalve 121 is in the closed state, the first channel 131 is in the closed state, and the first liquid cannot flow in the first channel 131.

[0103] In this embodiment, refer to Figure 4 When a single first target A and a single second target B are present in the paired flow channel 136, the first microvalve 121 and the second microvalve 122 are in the closed state (the microvalve is filled with black to indicate that it is in the closed state), and the paired microvalve 126 and the buffer microvalve 124 are in the open state (the microvalve is filled with gray to indicate that it is in the open state). At this time, the buffer in the buffer flow channel 134 can send the single first target A and the single second target B in the paired flow channel 136 into the oil in the oil flow channel 133 to form droplets C.

[0104] As described above, this embodiment has a separate pairing channel 136 for temporarily storing a single first target A and a single second target B, and a separate buffer solution channel 134. The buffer solution pushes the single first target A and the single second target B into the oil, eliminating the need for the first liquid and the second liquid to push them in. Therefore, the first channel 131 and the second channel 132 can remain in a closed state during the encapsulation process, thereby completely preventing other first targets and / or other second targets from entering the droplet C. This is beneficial for improving the success rate of encapsulation and reducing cell loss caused by multiple encapsulations.

[0105] Based on the first embodiment, referring to Figure 2 , Figure 3In some embodiments of the present invention, the microfluidic system further includes a waste liquid channel 135, which is used to discharge the first liquid and the second liquid, and is connected to a paired channel 136. Furthermore, the microfluidic system also includes a waste liquid microvalve 125 corresponding to the waste liquid channel 135, which is used to control the opening and closing of the waste liquid channel 135. When the paired microvalve 126 is closed and the first microvalve 121 and the waste liquid microvalve 125 are open, the first liquid in the first channel 131 can flow out through the paired channel 136 and the waste liquid channel 135. It should be noted that the first target object will gradually approach and reach the mating flow channel 136 following the flow of the first liquid. When the first target object has not yet reached the mating flow channel 136, the first flow channel 131 will continuously discharge the first liquid. This first liquid needs to be discharged as waste liquid. In this embodiment, after the first liquid is discharged from the first flow channel 131, it will first enter the mating flow channel 136, and then enter the waste liquid flow channel 135 for discharge. In other words, in this embodiment, the first liquid will not pass through the oil flow channel 133 during discharge. Figure 1 Compared to the proposed solution, this solution also has the following advantages:

[0106] 1. Figure 1 In the scheme shown, since the waste liquid needs to pass through the oil channel 3 for discharge, it will inevitably take away the oil in the oil channel 3, resulting in oil waste. However, in this embodiment, the first liquid and the second liquid do not pass through the oil channel 133 when they are discharged, so they will not take away the oil in the oil channel 133, thus reducing the amount of oil used.

[0107] 2. Figure 1 In the illustrated scheme, since waste liquid discharge needs to pass through oil channel 3, oil channel 3 needs to be closed during waste discharge. However, after pairing, the first channel 1, the second channel 2, and the oil channel 3 need to be opened simultaneously; otherwise, oil channel 3 will not be able to supply oil in time and droplets cannot be formed. Figure 1 The scheme shown places extremely high demands on the control of the microvalves corresponding to the first flow channel 1, the second flow channel 2, and the oil flow channel 3, which often leads to the failure of the target object encapsulation and further results in the loss of the target object. However, in this embodiment, the first liquid and the second liquid do not pass through the oil flow channel 133 when they are discharged. Therefore, the oil flow channel 133 can always be filled with oil. After the pairing is completed, it is only necessary to open the buffer microvalves 124 and the pairing microvalves 126. Even if the buffer microvalves 124 and the pairing microvalves 126 are not synchronized, it will not cause the encapsulation failure.

[0108] 3. Figure 1In the illustrated scheme, after the oil channel 3 is closed, some oil will remain inside. When the waste liquid passes through this residual oil, it will generate empty droplets. The empty droplets will cause the normal droplets to mix. Therefore, the empty droplets and the normal droplets need to be assisted by the micro valves of the waste liquid channel 4 and the sample channel 5 to enter different channels. However, this also places extremely high demands on the control of the two micro valves, otherwise it will lead to losses. For example, when droplets are formed, if the micro valve of the waste liquid channel 4 fails to close in time and the micro valve of the sample channel 5 fails to open in time, the successfully encapsulated droplets will also enter the waste liquid channel 4. In this embodiment, the first liquid and the second liquid will not pass through the oil channel 133 when they are discharged. The encapsulated droplet C can reach the subsequent storage position with the flow of oil in the oil channel 133 without involving the selection of subsequent channels. On the one hand, it reduces the control difficulty, and on the other hand, it avoids the loss of the target object due to this reason.

[0109] 4. As mentioned above, Figure 1 The proposed solution generates a large number of empty droplets, necessitating valve switching for sorting, which significantly impacts processing speed compared to... Figure 1 The solution avoids the generation of a large number of empty droplets in the microfluidic chip structure of this embodiment, and eliminates the need for sorting of empty droplets and sample droplets, thus improving the pairing efficiency. For example, it can generate 5 to 10 droplets per second, or 18,000 to 36,000 droplets per hour. In addition, the microfluidic chip structure of this embodiment can also significantly reduce the control difficulty of microvalves.

[0110] In some embodiments, the microfluidic system further includes a power element such as a pump for driving the flow of the buffer solution. In some specific embodiments, the microfluidic system includes a high-speed pump capable of achieving high-speed flow and stoppage of the buffer solution, thereby adapting to high-speed pairing.

[0111] Based on the first embodiment, referring to Figure 2 , Figure 3 In some embodiments of the present invention, the waste liquid channel 135 is directly connected to the paired channel 136. The microfluidic system also includes a waste liquid microvalve 125 corresponding to the waste liquid channel 135, which is used to control the opening and closing of the waste liquid channel 135. When the paired microvalve 126 is closed and the second microvalve 122 and the waste liquid microvalve 125 are open, the liquid in the second channel 132 can flow out through the paired channel 136 and the waste liquid channel 135. The discharge process of the second liquid can be understood with reference to the first liquid, and will not be described in detail here.

[0112] When the waste liquid flow channel 135 is directly connected to the paired flow channel 136, refer to Figure 2 , Figure 3In some embodiments of the present invention, the first flow channel 131, the second flow channel 132, the buffer solution flow channel 134, and the waste liquid flow channel 135 are all located on the side of the paired microvalve 126 away from the oil flow channel 133. In this way, the first liquid, the second liquid, etc., can be prevented from entering the oil flow channel 133 before the pairing is completed. Specifically, in the illustrated embodiment, the oil flow channel 133 is distributed in a horizontal direction, and the paired flow channel 136 is distributed in a vertical direction with its lower end connected to the oil flow channel 133. Thus, the paired microvalve 126 is approximately located at the lower end of the paired flow channel 136, and the first flow channel 131, the second flow channel 132, the buffer solution flow channel 134, and the waste liquid flow channel 135 are all located above the paired microvalve 126.

[0113] Based on the above structure, the control of each micro-valve is described as follows:

[0114] 1. Pairing process

[0115] The buffer solution microvalve 124 and its paired microvalve 126 are closed, while the first microvalve 121, the second microvalve 122, and the waste liquid microvalve 125 are opened, allowing the first and second liquids to drain through the paired flow channel 136 and the waste liquid flow channel 135. Upon detection of a single first target object, the first microvalve 121 closes, and the single first target object stops within the paired flow channel 136 (e.g., ...). Figure 4 (b) When a single second target is detected, the second microvalve 122 closes, and the single second target stops within the mating flow channel 136 (e.g., ...). Figure 4 (c) When both the first and second target objects stop within the paired flow channel 136, the waste liquid micro-valve 125 closes (e.g., Figure 4 (c in the text)

[0116] 2. Packaging process

[0117] When the first microvalve 121, the second microvalve 122, and the waste liquid microvalve 125 are closed, the buffer solution microvalve 124 and the paired microvalve 126 are opened. The buffer solution delivers a single first target and a single second target from the paired flow channel 136 into the oil in the oil flow channel 133 to form droplets (e.g., Figure 4 (d in the text)

[0118] When the waste liquid flow channel 135 is directly connected to the paired flow channel 136, refer to Figure 3 In some embodiments of the present invention, along the extending direction of the paired flow channel 136, the waste liquid flow channel 135 is located between the first flow channel 131 and the second flow channel 132. In this way, when the first liquid is discharged, it will not drive the already stationary single second target object B. Similarly, when the second liquid is discharged, it will not drive the already stationary single second target object A. The discharge of the first liquid and the second liquid can be carried out synchronously.

[0119] When the waste liquid flow channel 135 is directly connected to the paired flow channel 136, refer to Figure 3 In some embodiments of the present invention, the first flow channel 131 and the second flow channel 132 are located on the same side of the paired flow channel 136, and the waste liquid flow channel 135 is located on the other side of the paired flow channel 136. Specifically to Figure 3 In the embodiment shown, the first flow channel 131 and the second flow channel 132 are located on the left side of the paired flow channel 136, and the waste liquid flow channel 135 is located on the right side of the paired flow channel 136. In this way, the first liquid and the second liquid can flow in approximately the same direction, which facilitates the discharge of waste liquid.

[0120] When the waste liquid flow channel 135 is directly connected to the paired flow channel 136, refer to Figure 3 In this embodiment of the invention, at least a first portion 1311 of the first flow channel 131 that communicates with the paired flow channel 136 is intersected with the paired flow channel 136, specifically referring to... Figure 3 In the embodiment shown, the first portion 1311 is perpendicular to the mating channel 136.

[0121] When the waste liquid flow channel 135 is directly connected to the paired flow channel 136, refer to Figure 3 In this embodiment of the invention, the second portion 1321 of the second flow channel 132, which is at least connected to the paired flow channel 136, is intersected with the paired flow channel 136, specifically referring to... Figure 3 In the embodiment shown, the second portion 1321 is perpendicular to the mating channel 136.

[0122] When the waste liquid flow channel 135 is directly connected to the paired flow channel 136, refer to Figure 3 The third part 1351 of the waste liquid flow channel 135, which is at least connected to the paired flow channel 136, is intersected with the paired flow channel 136, as specifically referred to Figure 3 In the embodiment shown, the third portion 1351 is perpendicular to the mating channel 136.

[0123] When the waste liquid flow channel 135 is directly connected to the paired flow channel 136, refer to Figure 3 The fourth portion 1341 of the buffer solution channel 134, which is at least connected to the paired channel 136, is coaxially arranged with the paired channel 136. Thus, the buffer solution can enter the paired channel 136 in the same direction and carry the single first target A and the single second target B into the oil. It should be noted that in this embodiment, the buffer solution channel 134 and the paired channel 136 can be two parts of a single channel. Figure 3 As shown in the figure, the lower part of the vertical straight channel is the pairing channel 136, and the upper part is the buffer channel 134. For ease of reading, the range of the pairing channel 136 is roughly marked by a dashed box in the figure. However, the dashed box should not be interpreted as a specific limitation on the shape and length of the pairing channel 136.

[0124] It is understood that the above embodiments can be combined, that is, the first part 1311, the second part 1321 and the third part 1351 are all intersecting with the mating channel 136, while the fourth part 1341 is coaxially arranged with the mating channel 136.

[0125] Based on the first embodiment, some embodiments of the microfluidic system of the present invention further include a detection module. The detection module is configured to identify a single first target A within the first flow channel 131. When the detection module identifies the single first target A, both the first microvalve 121 and the paired microvalve 126 are in a closed state to keep the single first target A within the paired flow channel 136. It should be noted that in this embodiment, the first microvalve 121 being in a closed state specifically means switching from an open state to a closed state, and the paired microvalve 126 being in a closed state specifically means remaining in a closed state.

[0126] In other embodiments, the detection module is configured to identify a single second target object B within the second flow channel 132. When the detection module identifies the single second target object B, both the second microvalve 122 and the paired microvalve 126 are in a closed state to keep the single second target object B within the paired flow channel 136. It should be noted that in this embodiment, the second microvalve 122 being in a closed state specifically means switching from an open state to a closed state, and the paired microvalve 126 being in a closed state specifically means remaining in a closed state.

[0127] When a single first target object A is identified by the detection module, in some embodiments of the present invention, the microfluidic system further includes a waste liquid microvalve 125 corresponding to the waste liquid flow channel 135. The waste liquid microvalve 125 is used to control the opening and closing of the waste liquid flow channel 135. Specifically, when the detection module does not identify a single first target object A, the paired microvalve 126 is in a closed state, and the first microvalve 121 and the waste liquid microvalve 125 are in an open state, allowing the liquid in the first flow channel 131 to flow out through the paired flow channel 136 and the waste liquid flow channel 135. It should be noted that in this embodiment, the paired microvalve 126 being in a closed state specifically means remaining in a closed state, and the first microvalve 121 and the waste liquid microvalve 125 being in an open state specifically means remaining in an open state.

[0128] In some embodiments of the present invention, when the detection module identifies a single second target object B, if the detection module does not identify a single second target object B, the paired microvalve 126 is in a closed state while the second microvalve 122 and the waste liquid microvalve 125 are in an open state, so that the liquid in the second flow channel 132 can flow out through the paired flow channel 136 and the waste liquid flow channel 135. It should be noted that in this embodiment, the paired microvalve 126 being in a closed state specifically means being kept in a closed state, and the second microvalve 122 and the waste liquid microvalve 125 being in an open state specifically means being kept in an open state.

[0129] When a single first target object A is identified by the detection module, in some embodiments of the present invention, the detection module is specifically a visual detection module, which includes a camera and a controller. The controller is configured to control the camera to capture images. In this embodiment, the image captured by the camera includes at least a first image of the first portion 1311 where the first flow channel 131 and the pairing flow channel 136 communicate. After the camera captures the image, the controller is also configured to identify the single first target object A based on the first image. In other words, the single first target object A has already completed the identification operation before entering the pairing flow channel 136. It should be noted that the microspheres in this embodiment can be magnetic microspheres, which can be adsorbed by magnetic components for recycling.

[0130] In other embodiments, the image captured by the camera includes at least a second image of the second portion 1321 in which the second flow channel 132 communicates with the pairing flow channel 136, and the controller is also configured to identify a single second target object B based on the second image. In other words, the single second target object B has already completed the identification operation before entering the pairing flow channel 136.

[0131] In the above process, after the image captured by the camera is transmitted to the controller, the controller detects the target in the detection area (e.g., an area with a length and width of 200 pixels), classifies the cropped target image through a model trained by a convolutional neural network, and judges and controls the corresponding micro valve based on the classification output.

[0132] When the detection module identifies a single first target A through an image, in some embodiments of the present invention, the image captured by the camera also includes at least a third image of the pairing channel 136. The controller is configured to identify whether a single first target A exists in the pairing channel 136 based on the third image. In other words, in this embodiment, the image captured by the camera can also be used to verify whether a single first target A has entered the pairing channel 136.

[0133] In other embodiments, the images captured by the camera also include at least a third image of the pairing channel 136, and the controller is configured to identify whether a single second target object B exists in the pairing channel 136 based on the third image. In other words, in this embodiment, the images captured by the camera can also be used to verify whether a single second target object B has entered the pairing channel 136.

[0134] It should be noted that the above embodiments can be combined. For example, the images captured by the camera include the aforementioned first image, second image, and third image. In this way, the controller can perform combined operations through the same image. For example, it can simultaneously identify a single first target A and a single first target B, or identify a single first target A and determine whether a single second target B has reached the pairing channel 136, or identify a single first target B and determine whether a single first target A has reached the pairing channel 136, or determine whether a single first target A and a single second target B have reached the pairing channel 136.

[0135] When a single first target object A is identified by the detection module, in some embodiments of the present invention, the detection module is specifically a fluorescence detection module, which includes a first light source, a first optical detection device, and a controller. The controller is configured to control the first light source to emit a first detection light 210 into the first flow channel 131. The first detection light 210 can excite a first fluorescence after irradiating the single first target object A. The first optical detection device can receive the first fluorescence excited by the single first target object A. For example, the first light source is a laser, and the first optical detection device is a photomultiplier tube. After receiving the first fluorescence emitted by the single first target object A, the photomultiplier tube can convert the light signal into an electrical signal and transmit it to the controller. It should be noted that the microspheres in this embodiment can be fluorescent magnetic microspheres stained with fluorescent substances, which can emit fluorescence under the excitation of an external light source.

[0136] Among them, reference Figure 4 The first light source projects the first detection light 210 onto the set position of the first flow channel 131. When no cell reaches the set position, the first light detection device will not receive the light signal. When a cell reaches the set position, the cell is excited by the first detection light 210 and produces fluorescence. When the first light detection device detects the first fluorescence, the detection device identifies a single first target A.

[0137] In some embodiments of the present invention, when a single second target object B is identified by the detection module, the detection module is specifically a fluorescence detection module, a second light source, a second light detection device, and a controller. The controller is configured to control the second light source to emit a second detection light 220 into the second flow channel 132. The second detection light 220 can excite a second fluorescence after irradiating the single second target object B. When the second light detection device detects the second fluorescence, the controller identifies the single second target object B.

[0138] Combination Figure 4 Describe a fluorescence-based identification scheme for first target A / second target B:

[0139] 1. Identification of the first target object A

[0140] When the first microvalve 121 is in the open state, the first liquid continuously flows through the first detection position irradiated by the first detection light 210. When a single first target A passes through the first detection position with the first liquid, it is excited by the first detection light 210 to generate first fluorescence. After the first fluorescence is received by the first optical detection device, an electrical signal is generated. The controller identifies the single first target A based on the electrical signal and then controls the first microvalve 121 to switch to the closed state, so that the single first target A stops in the paired flow channel 136.

[0141] 2. Second target object identification

[0142] When the second microvalve 122 is in the open state, the second liquid continuously flows through the second detection position irradiated by the second detection light 220. When a single second target object B passes through the second detection position with the second liquid, it is excited by the second detection light 220 to generate second fluorescence. After the second fluorescence is received by the second light detection device, an electrical signal is generated. The controller identifies the single second target object B based on the electrical signal and then controls the second microvalve 122 to switch to the closed state, so that the single second target object B stops in the paired flow channel 136.

[0143] It should be noted that the first light source and the second light source can be lasers of different wavelengths.

[0144] When a single first target object A is identified by the detection module, in some embodiments of the present invention, the detection module is specifically an electrode detection module, see reference. Figure 5 The system includes a first detection electrode 230 and a controller. The first detection electrode 230 extends into a first flow channel 131. When a single first target object A passes through the first detection electrode 230, the first detection electrode 230 generates a corresponding signal. The controller is configured to identify the single first target object A based on the signal detected by the first detection electrode 230. Specifically, the first detection electrode 230 includes a positive electrode and a negative electrode, which are arranged side by side. When the microfluidic system includes a microfluidic chip 100, the first detection electrode 230 can be a metal layer disposed between the control layer 120 and the flow channel layer 130 of the microfluidic chip 100. Furthermore, in this embodiment, the signal detected by the first detection electrode 230 can be a change in amplitude and phase difference. It should be noted that the microspheres in this embodiment can be magnetic microspheres, which can be adsorbed by magnetic components for recycling.

[0145] When a single second target object B is identified by the detection module, in some embodiments of the present invention, the detection module is specifically an electrode detection module, see reference. Figure 5 It includes a second detection electrode 240 and a controller. The second detection electrode 240 extends into the second flow channel 132, and the controller is configured to identify a single second target object B based on the signal detected by the second detection electrode 240. The second detection electrode 240 can be understood with reference to the first detection electrode 230.

[0146] In some embodiments of the present invention, when a single first target object A is identified by the detection module, after the detection module identifies the single first target object A, the first microvalve 121 switches from an open state to a closed state after a set delay, so that the single first target object A enters the pairing flow channel 136 from the first flow channel 131. (Refer to...) Figure 4 , Figure 5 Since the identification position of a single first target object A is at a certain distance from the pairing flow channel 136, this embodiment uses a delayed closing method for the first micro-valve 121 to ensure that the single first target object A can enter the pairing flow channel 136. It should be noted that the set time of the delay can be determined according to the distance between the identification position and the pairing flow channel 136 and the flow rate of the first liquid.

[0147] In some embodiments of the present invention, when a single second target object B is identified by the detection module, after the detection module identifies the single second target object B, the second microvalve 122 switches from an open state to a closed state after a set delay, so that the single second target object B enters the pairing flow channel 136 from the second flow channel 132. (Refer to...) Figure 4 , Figure 5 Since the identification position of a single second target object B is at a certain distance from the pairing flow channel 136, this embodiment uses a delayed closing method for the second micro-valve 122 to ensure that the single second target object B can enter the pairing flow channel 136. It should be noted that the set time of the delay can be determined according to the distance between the identification position and the pairing flow channel 136 and the flow rate of the second liquid.

[0148] Based on the first embodiment, referring to Figure 6 , Figure 7 In some embodiments of the present invention, the microfluidic system further includes a liquid injection system for injecting liquid, such as reverse transcription fluid, into the droplet C. Specifically, the microfluidic system further includes a liquid injection channel 137, which is connected to an oil channel 133. The connection between the paired channel 136 and the oil channel 133 is defined as a first connection, and the connection between the liquid injection channel 137 and the oil channel 133 is defined as a second connection. The second connection is located downstream of the first connection along the flow direction of the oil in the oil channel 133.

[0149] The microfluidic system also includes a fusion electrode 300, which applies an electric field to the second connecting portion. When droplet C passes through the second connecting portion, the liquid in the injection channel 137 fuses into droplet C under the influence of the electric field. The liquid in the injection channel 137 is also insoluble in the oil, and the pressure within the injection channel 137 is relatively low, causing the liquid in the injection channel 137 to slightly protrude into the oil channel 133. Thus, when droplet C passes through the second connecting portion, it can contact the liquid in the injection channel 137, facilitating their fusion.

[0150] When the microfluidic system also includes a liquid injection system, refer to Figure 6 , Figure 7 In some embodiments of the present invention, the flow channel layer 130 has a processing area, within which the first flow channel 131, the second flow channel 132, the oil flow channel 133, the buffer flow channel 134, the waste liquid flow channel 135, the paired flow channel 136, and the injection flow channel 137 are all located. It should be noted that the processing area is for illustrative purposes and does not imply that the microfluidic system must have a clearly defined and identifiable region.

[0151] In this embodiment, the fusion electrode 300 includes a first electrode 310 and a second electrode 320, wherein the first electrode 310 is arranged around the processing area to achieve a better electrofusion effect. In some specific embodiments, when the microfluidic system includes a microfluidic chip 100, an electrode channel is formed on the flow channel layer 130 of the microfluidic chip 100, and a metal rod is inserted into the electrode channel. When the metal rod is heated, the metal rod melts and fills the electrode channel, and after cooling, the first electrode 310 and the second electrode 320 are formed. In some embodiments, the electrode channel has an electrode injection port to allow the metal rod to be injected, so as to achieve a better electrofusion effect. Figure 6 As shown in the example, the electrode injection port includes a first injection port 1391 and a second injection port 1392 at both ends of the first electrode channel, and a third injection port 1393 at one end of the second electrode channel. It should be noted that the length of the first electrode channel is relatively long in this embodiment. When the molten metal fills the electrode channel, the other injection port can be used as an exhaust port, so that the molten metal can fill the entire electrode channel.

[0152] When the fusion electrode 300 includes a first electrode 310 and a second electrode 320, and the first electrode 310 surrounds the processing area, refer to Figure 7 In some embodiments of the present invention, the two ends of the first electrode 310 are arranged side by side with the second electrode 320, and the second electrode 320 is arranged between the two ends of the first electrode 310 to achieve a better electrofusion effect.

[0153] Based on the first embodiment, referring to Figure 2In some embodiments of the present invention, the microfluidic chip 100 further includes an oil microvalve 123 corresponding to the oil flow channel 133. The oil microvalve 123 is used to control the opening and closing of the oil flow channel 133. When the oil microvalve 123 is open, the oil in the oil flow channel 133 flows, driving the droplet C to move downstream. Specifically, in the illustrated embodiment, the oil microvalve 123 is located upstream of the first connecting portion along the flow direction of the oil in the oil flow channel 133.

[0154] Based on the first embodiment, in some embodiments of the present invention, the microfluidic system further includes a first microvalve control channel 138, as shown in the reference. Figure 8 The first microvalve 121 includes a diaphragm 1211 disposed between the first flow channel 131 and the first microvalve control flow channel 138. When the first microvalve 121 is in such a state... Figure 8 When in the open state shown by a, the diaphragm 1211 is in a horizontal state, and the first liquid can flow within the first flow channel 131; when the diaphragm 1211 is driven to bulge into the first flow channel 131 to be in the open state as shown by a, the diaphragm 1211 is in a horizontal state, and the first liquid can flow within the first flow channel 131; when the diaphragm 1211 is driven Figure 8 When in the closed state as shown in b, the diaphragm 1211 is close to the inner wall of the first flow channel 131 to block the first flow channel 131, and the first liquid cannot flow in the first flow channel 131.

[0155] In this embodiment, the inner wall of the first flow channel 131 is an arc-shaped inner wall, which facilitates the bonding with the diaphragm 1211 protruding into the first flow channel 131, thereby ensuring the cut-off effect.

[0156] It should be noted that, in addition to the aforementioned first microvalve control channel 138, the microfluidic system also includes microvalve control channels corresponding to the second microvalve 122, oil microvalve 123, buffer microvalve 124, waste liquid microvalve 125, and paired microvalve 126. Accordingly, the second microvalve 122, oil microvalve 123, buffer microvalve 124, waste liquid microvalve 125, and paired microvalve 126 all include diaphragms. Their specific settings and control methods can be understood by referring to the first microvalve control channel 138, and will not be described in detail here.

[0157] It should also be noted that the power to drive the diaphragm 1211 to bulge can be air pressure. In this embodiment, the first microvalve control channel 138 is filled with liquid. An external air source drives the liquid in the first microvalve control channel 138 to flow, thereby pushing the diaphragm 1211 to bulge or reset. Compared with the method of direct air pressure drive, since liquid is difficult to compress, this method can achieve rapid response of the diaphragm 1211, thereby adapting to high-speed pairing.

[0158] Based on the first embodiment, referring to Figure 9In some embodiments of the present invention, the microfluidic system includes a microfluidic chip 100, which includes a base layer 110, a control layer 120 and a flow channel layer 130 stacked sequentially. The base layer 110 may be made of glass, and the control layer 120 and the flow channel layer 130 may be made of polydimethylsiloxane (PDMS).

[0159] The flow channel layer 130 includes the aforementioned first flow channel 131, second flow channel 132, oil flow channel 133, buffer flow channel 134, waste liquid flow channel 135, and paired flow channel 136. The control layer 120 includes the aforementioned paired microvalve 126, first microvalve 121, second microvalve 122, and buffer microvalve 124, as well as microvalve control flow channels corresponding to each microvalve.

[0160] Based on the first embodiment, in some embodiments of the present invention, the first target A is a cell, and the second target B is a microsphere. In some embodiments, the microspheres can be further configured as magnetic microspheres, so that the RNA released after cell lysis can be adsorbed onto the surface of the microspheres, and the microspheres can be collected subsequently by a magnetic component, thereby collecting the RNA. In some embodiments, the microspheres can be further configured as fluorescent magnetic microspheres so that they emit fluorescence when excited by external light.

[0161] Based on the first embodiment, in some embodiments of the present invention, the first target A is a first cell and the second target B is a second cell, used to study the interaction effects between cells.

[0162] The second embodiment of the present invention proposes a microfluidic system for target pairing, which differs from the first embodiment in that: in the first embodiment, the first target A and the second target B of the microfluidic system stop in the pairing channel 136, while in this embodiment, the first target A and the second target B stop in the first channel 131 and the second channel 132.

[0163] Reference Figure 10The microfluidic system includes a first channel 131, a second channel 132, an oil channel 133, a buffer channel 134, and a paired channel 136. The first channel 131 is for a first liquid containing a first target substance to pass through; the second channel 132 is for a second liquid containing a second target substance to pass through; the buffer channel 134 is for a buffer solution to pass through; and the oil channel 133 is for an oil solution that is immiscible with the first liquid, the second liquid, and the buffer solution to pass through. The paired channel 136 is connected to the oil channel 133. Both the first channel 131 and the second channel 132 are connected to the paired channel 136. The buffer channel 134 is connected to both the first channel 131 and the second channel 132. The first channel 131 has a first portion 1311 connected to the paired channel, and the second channel 132 has a second portion 1321 connected to the paired channel. In some further embodiments, the microfluidic system also includes a waste liquid channel 135 for discharging the first liquid and the second liquid.

[0164] The microfluidic system of this embodiment also includes multiple microvalves, specifically including a first microvalves 121 corresponding to the first flow channel 131, a second microvalves 122 corresponding to the second flow channel 132, and a buffer microvalves 124 corresponding to the buffer flow channel 134. The first microvalves 121 are used to control the opening and closing of the first flow channel 131, the second microvalves 122 are used to control the opening and closing of the second flow channel 132, and the buffer microvalves 124 are used to control the opening and closing of the buffer flow channel 134. The opening and closing here can be understood with reference to the first embodiment.

[0165] In this embodiment, refer to Figure 10 When a single first target A is present in the first part 1311 and a single second target B is present in the second part 1321, the first microvalve 121 and the second microvalve 122 are closed, and the buffer microvalve 124 is open, so that the buffer in the buffer channel 134 first sends the single first target A in the first part 1311 and the single second target B in the second part 1321 into the paired channel 136, and then together into the oil in the oil channel 133 to form droplets.

[0166] As described above, this embodiment has a first part 1311 of the first flow channel 131 for temporarily storing a single first target A, a second part 1321 of the second flow channel 132 for temporarily storing a single second target B, and a separate buffer flow channel 134. The buffer solution pushes the single first target A and the single second target B into the oil, eliminating the need for the first liquid and the second liquid to push them. Therefore, the first flow channel 131 and the second flow channel 132 can remain in a closed state during the encapsulation process, thereby completely preventing other first targets and / or other second targets from entering the droplet C. This is beneficial for improving the success rate of encapsulation and reducing cell loss caused by multiple encapsulations.

[0167] Based on the second embodiment, referring to Figure 10 In some embodiments of the present invention, the microfluidic system further includes a waste liquid channel 135 and a droplet channel 139. The waste liquid channel 135 is used to discharge excess first liquid and second liquid, and the droplet channel 139 is used to discharge droplets C. Both the waste liquid channel 135 and the droplet channel 139 are connected to the oil channel 133. Furthermore, the microfluidic system also includes a waste liquid microvalve 125 corresponding to the waste liquid channel 135 and a droplet microvalve 127 corresponding to the droplet channel 139. The waste liquid microvalve 125 is used to control the opening and closing of the waste liquid channel 135, and the droplet microvalve 127 is used to control the opening and closing of the droplet channel 139. When the first microvalve 121 and the waste liquid microvalve 125 are in the open state, and the droplet microvalve 127 is in the closed state, the first liquid in the first channel 131 can flow out through the paired channel 136, the oil channel 133, and the waste liquid channel 135.

[0168] Based on the second embodiment, referring to Figure 10 In some embodiments of the present invention, the microfluidic system further includes a waste liquid channel 135, a droplet channel 139, a waste liquid microvalve 125 corresponding to the waste liquid channel 135, and a droplet microvalve 127 corresponding to the droplet channel 139. The waste liquid channel 135, droplet channel 139, waste liquid microvalve 125, and droplet channel 139 can all be understood with reference to the foregoing embodiments. When the second microvalve 122 and the waste liquid microvalve 125 are in the open state, and the droplet microvalve 127 is in the closed state, the second liquid in the second channel 132 can flow out through the paired channel 136, the oil channel 133, and the waste liquid channel 135. The discharge process of the second liquid can be understood with reference to the first liquid, and will not be described in detail here.

[0169] Based on the second embodiment, referring to Figure 10In some embodiments of the present invention, the waste liquid channel 135, the droplet channel 139, the waste liquid micro-valve 125 corresponding to the waste liquid channel 135, and the droplet micro-valve 127 corresponding to the droplet channel 139 can all be understood with reference to the foregoing embodiments. In this embodiment, when droplets are formed in the oil in the oil channel 133, the waste liquid micro-valve 125 is in a closed state, and the droplet micro-valve 127 is in an open state, so that the droplets are discharged through the droplet channel 139. In conjunction with the foregoing embodiments, the switching between waste liquid discharge and droplet discharge can be achieved by switching between the waste liquid micro-valve 125 and the droplet channel 139.

[0170] Based on the second embodiment, referring to Figure 10 In some embodiments of the present invention, the microfluidic system further includes a waste liquid channel 135, a droplet channel 139, a waste liquid microvalve 125 corresponding to the waste liquid channel 135, and a droplet microvalve 127 corresponding to the droplet channel 139. The paired channel 136 is connected to the inlet end of the oil channel 133, and the waste liquid channel 135 and the droplet channel 139 are connected to the outlet end of the oil channel 133. Specifically... Figure 10 In the illustrated embodiment, the paired flow channel 136 is connected to the upper end of the oil flow channel 133, and the waste liquid flow channel 135 and the droplet flow channel 139 are connected to the lower end of the oil flow channel 133. Furthermore, the first flow channel 131 and the second flow channel 132 are both connected to the inlet end of the paired flow channel 136. The buffer solution flow channel 134 has two branch channels at its end, which are respectively connected to the first portion 1311 and the second portion 1321, and the two branch channels are of equal length. Thus, the buffer solution allows the first target A in the first portion 1311 and the second target B in the second portion 1321 to be fed into the paired flow channel 136 approximately synchronously.

[0171] Based on the second embodiment, referring to Figure 10 In some embodiments of the present invention, the microfluidic system further includes an oil microvalve 123 corresponding to the oil flow channel 133. When draining waste liquid, the oil microvalve 123 is in a closed state, and the waste liquid can enter the waste liquid flow channel 135 through the oil flow channel 133. When encapsulating, the oil microvalve 123 is in an open state, and the oil flow channel 133 will be filled with oil.

[0172] Based on the second embodiment, referring to Figure 10In some embodiments of the present invention, the microfluidic system further includes a detection module configured to identify a single first target object within the first flow channel 131. When the detection module detects a single first target object, the first microvalve 121 is in a closed state to keep the single first target object within the first portion 1311. It should be noted that in this embodiment, the first microvalve 121 being in a closed state specifically refers to switching from an open state to a closed state.

[0173] In other embodiments, the detection module is configured to identify a single second target object within the second flow channel 132, wherein, upon detection of the single second target object, the second microvalve 122 is in a closed state to retain the single second target object within the second portion 1321. It should be noted that, in this embodiment, the second microvalve 122 being in a closed state specifically refers to switching from an open state to a closed state.

[0174] When a single first target object A is identified by the detection module, in some embodiments of the present invention, the microfluidic system further includes a waste liquid microvalve 125 corresponding to the waste liquid channel 135, which is used to control the opening and closing of the waste liquid channel 135. Specifically, when the detection module does not identify a single first target object A, the droplet microvalve 127 is in a closed state, while the first microvalve 121 and the waste liquid microvalve 125 are in an open state, allowing the liquid in the first channel 131 to flow out through the paired channel 136, the oil channel 133, and the waste liquid channel 135. It should be noted that in this embodiment, the droplet microvalve 127 being in a closed state specifically means remaining in a closed state, and the first microvalve 121 and the waste liquid microvalve 125 being in an open state specifically means remaining in an open state.

[0175] In some embodiments of the present invention, when the detection module identifies a single second target object B, if the detection module does not identify a single second target object B, the droplet microvalve 127 is in a closed state while the second microvalve 122 and the waste liquid microvalve 125 are in an open state, so that the liquid in the second flow channel 132 can flow out through the paired flow channel 136, the oil flow channel 133, and the waste liquid flow channel 135. It should be noted that in this embodiment, the droplet microvalve 127 being in a closed state specifically means being kept in a closed state, and the second microvalve 122 and the waste liquid microvalve 125 being in an open state specifically means being kept in an open state.

[0176] The different detection methods of the detection module can be understood by referring to the visual detection, fluorescence detection and electrode detection in the first embodiment, and will not be described in detail here.

[0177] The third embodiment of the present invention also proposes a pairing method, comprising the following steps:

[0178] S100 Identifies a single first target object from the first liquid carrying the first target object, and stops the first liquid after identifying the single first target object so that the single first target object stops in the target area. In some embodiments, this step can be performed based on the microfluidic system of the first embodiment or the second embodiment described above, that is, the target area in this step can be the paired flow channel 136 in the first embodiment or the first part 1311 in the second embodiment.

[0179] S200 Identifies a single second target object from the second liquid carrying the second target object, and stops the second liquid after identifying the single second target object so that the single second target object stops in the target area. In some embodiments, this step can be performed based on the microfluidic system of the first embodiment or the second embodiment described above, that is, the target area in this step can be the paired flow channel 136 in the first embodiment or the second part 1321 in the second embodiment.

[0180] S300 When both the first target object and the second target object stop in the target area, and the first liquid and the second liquid are in a stopped state, the third liquid sends the first target object and the second target object in the target area into the oil to form droplets.

[0181] It should be noted that there is no restriction on the order of steps S100 and S200.

[0182] Based on the third embodiment, in some embodiments of the present invention, the method for identifying a single first target object from a first liquid carrying a first target object includes the following steps: acquiring an image of a detection area, identifying a single first target object based on image information within the image, wherein the first liquid is able to enter the target area after passing through the detection area.

[0183] Based on the third embodiment, in some embodiments of the present invention, the method for identifying a single second target object from a second liquid carrying a second target object includes the following steps: acquiring an image of a detection area, identifying a single second target object based on image information within the image, wherein the second liquid is able to enter the target area after passing through the detection area.

[0184] The foregoing embodiments can be understood with reference to the visual detection scheme in the first embodiment.

[0185] Based on the third embodiment, in some embodiments of the present invention, a method for identifying a single first target object from a first liquid carrying a first target object includes the following steps: detecting a light signal within a detection area, and identifying a single first target object based on the detected light signal, wherein the first liquid can enter the target area after passing through the detection area. The light signal may be fluorescence generated by the first target object upon stimulation.

[0186] Based on the third embodiment, in some embodiments of the present invention, a method for identifying a single second target object from a second liquid carrying a second target object includes the following steps: detecting a light signal within a detection area, and identifying a single second target object based on the detected light signal, wherein the second liquid is capable of entering a target area after passing through the detection area. The light signal may be fluorescence generated by the second target object upon stimulation.

[0187] The foregoing embodiments can be understood with reference to the fluorescence detection scheme in the first embodiment.

[0188] Based on the third embodiment, in some embodiments of the present invention, the method for identifying a single first target object from a first liquid carrying a first target object includes the following steps: detecting an electrical signal generated by an electrode disposed in a detection area, and identifying a single first target object based on the detected electrical signal, wherein the first liquid can enter the target area after passing through the detection area.

[0189] Based on the third embodiment, some embodiments of the present invention include a method for identifying a single second target object from a second liquid carrying a second target object, comprising the following steps: detecting an electrical signal generated by an electrode disposed in a detection area, and identifying a single second target object based on the detected electrical signal, wherein the second liquid is capable of entering a target area after passing through the detection area. In this embodiment, the signal detected by the electrode may be a change in amplitude and phase difference.

[0190] In the aforementioned embodiments, the signal detected by the electrode can be a change in amplitude and phase difference.

[0191] The foregoing embodiments can be understood with reference to the electrode detection scheme in the first embodiment.

[0192] In the foregoing embodiments, the detection area is located upstream of the target area along the flow direction of the first liquid. For example, when the target area is the paired flow channel 136 in the first embodiment, the detection area can be the first part 1311 in the first flow channel 131 that connects to the paired flow channel 136. When the target area is the first part 1311 in the second embodiment, the detection area can be other areas in the first flow channel 131.

[0193] Along the flow direction of the second liquid, the detection area is located upstream of the target area. For example, when the target area is the paired flow channel 136 in the first embodiment, the detection area can be the second part 1321 in the second flow channel 132 that is connected to the paired flow channel 136. When the target area is the second part 1321 in the second embodiment, the detection area can be other areas in the second flow channel 132.

[0194] Based on the third embodiment, in some embodiments of the present invention, the pairing method further includes the following steps: allowing a droplet containing a single first target and a single second target to pass through a fusion region, and applying an electric field to the fusion region to incorporate a fourth liquid into the droplet. In this embodiment, the electric field can be applied through a fusion electrode.

[0195] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments, and various changes can be made within the scope of knowledge possessed by those skilled in the art without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof can be combined with each other unless otherwise specified.

Claims

1. A microfluidic system for target pairing, characterized in that, The microfluidic system includes a first channel, a second channel, an oil channel, a buffer channel, a waste channel, and a paired channel. The paired channel is connected to the oil channel. The first channel, the second channel, the waste channel, and the buffer channel are all connected to the paired channel. The first channel is used to allow a first liquid containing a first target to pass through. The second channel is used to allow a second liquid containing a second target to pass through. The first channel, the second channel, the buffer channel, and the waste channel are all located on the side of the paired microvalve away from the oil channel. The microfluidic system further includes a pairing microvalve corresponding to the paired flow channel, a first microvalve corresponding to the first flow channel, a second microvalve corresponding to the second flow channel, a waste liquid microvalve corresponding to the waste liquid flow channel, and a buffer liquid microvalve corresponding to the buffer liquid 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 flow channel, the second microvalve is used to control the opening and closing of the second flow channel, the buffer liquid microvalve is used to control the opening and closing of the buffer liquid flow channel, and the waste liquid microvalve is used to control the opening and closing of the waste liquid flow channel. When a single first target and a single second target exist within 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 delivers the single first target and the single second target in the paired flow channel into the oil in the oil flow channel to form droplets.

2. The microfluidic system for target pairing according to claim 1, characterized in that, in, When the paired microvalve is in the closed state and the first microvalve and the waste liquid microvalve are in the open state, the liquid in the first flow channel can flow out through the paired flow channel and the waste liquid flow channel; And / or, when the paired microvalve is in the closed state and the second microvalve and the waste liquid microvalve are in the open state, the liquid in the second flow channel can flow out through the paired flow channel and the waste liquid flow channel.

3. The microfluidic system for target pairing according to claim 2, characterized in that, The first flow channel and the second flow channel are located on the same side of the paired flow channels, and the waste liquid flow channel is located on the other side of the paired flow channels.

4. The microfluidic system for target pairing according to claim 2, characterized in that, The microfluidic system also includes at least one of the following: At least a first portion of the first flow channel that communicates with the paired flow channel is intersected with the paired flow channel; The second portion of the second flow channel, which is at least connected to the paired flow channel, is intersecting with the paired flow channel; At least a third portion of the waste liquid flow channel that is connected to the paired flow channel is intersected with the paired flow channel.

5. The microfluidic system for target pairing according to claim 2, characterized in that, The fourth portion of the buffer solution channel, which is at least connected to the paired channel, is coaxially arranged with the paired channel.

6. The microfluidic system for target pairing according to claim 2, characterized in that, Along the extension direction of the paired flow channels, the waste liquid flow channel is located between the first flow channel and the second flow channel.

7. The microfluidic system for target pairing according to claim 1, characterized in that, The microfluidic system further includes a detection module configured to identify a single first target object within the first flow channel. When the detection module identifies a single first target object, both the first microvalve and the paired microvalve are closed to keep the single first target object within the paired flow channel. And / or, the microfluidic system further includes a detection module configured to identify a single second target object within the second flow channel, wherein, when the detection module identifies a single second target object, both the second microvalve and the paired microvalve are closed to keep the single second target object within the paired flow channel.

8. The microfluidic system for target pairing according to claim 7, characterized in that, The microfluidic system further includes a waste liquid channel and a waste liquid microvalve corresponding to the waste liquid channel. The waste liquid channel is connected to the paired channel, and the waste liquid microvalve is used to control the opening and closing of the waste liquid channel. When the detection module fails to identify a single first target object, the paired microvalve is in a closed state and the first microvalve and the waste liquid microvalve are in an open state, so that the liquid in the first flow channel can flow out through the paired flow channel and the waste liquid flow channel; And / or, when the detection module does not identify a single second target, the paired microvalve is in a closed state and the second microvalve and the waste liquid microvalve are in an open state, so that the liquid in the second flow channel can flow out through the paired flow channel and the waste liquid flow channel.

9. The microfluidic system for target pairing according to claim 7, characterized in that, The detection module includes a camera and a controller, wherein the controller is configured to control the camera to capture images; The image includes at least a first image of a first portion of the first flow channel communicating with the paired flow channel, and the controller is further configured to identify the single first target object based on the first image; And / or, the image includes at least a second image of a second portion of the second flow channel communicating with the paired flow channel, and the controller is further configured to identify the single second target based on the second image.

10. The microfluidic system for target pairing according to claim 9, characterized in that, The image includes at least a third image of the paired flow channel, and the controller is configured to identify, based on the third image, whether the single first target and / or the single second target exists within the paired flow channel.

11. The microfluidic system for target pairing according to claim 7, characterized in that, The detection module includes a first light source, a first light detection device, and a controller. The controller is configured to control the first light source to emit a first detection light into the first flow channel. The first detection light can excite a first fluorescence after irradiating the single first target object. When the first light detection device detects the first fluorescence, the detection device identifies the single first target object. And / or, the detection module includes a second light source, a second light detection device, and a controller, wherein the controller is configured to control the second light source to emit a second detection light into the second flow channel, the second detection light being able to excite a second fluorescence after irradiating the single second target object, and the controller recognizing the single second target object when the second light detection device detects the second fluorescence.

12. The microfluidic system for target pairing according to claim 7, characterized in that, The detection module includes a first detection electrode and a controller. The first detection electrode extends into the first flow channel, and the controller is configured to identify the single first target object based on the signal detected by the first detection electrode. And / or, the detection module includes a second detection electrode and a controller, the second detection electrode extending into the second flow channel, and the controller being configured to identify the single second target object based on the signal detected by the second detection electrode.

13. The microfluidic system for target pairing according to claim 7, characterized in that, When the detection module detects the single first target object, the first micro valve switches from the open state to the closed state after a set delay, so that the single first target object enters the paired flow channel from the first flow channel. And / or, when the detection module detects the single second target object, the second microvalve switches from the open state to the closed state after a set delay, so that the single second target object enters the paired flow channel from the second flow channel.

14. The microfluidic system for target pairing according to claim 1, characterized in that, The microfluidic system further includes a liquid injection channel, which is connected to the oil channel. The connection between the paired channel and the oil channel is defined as the first connection point, and the connection between the liquid injection channel and the oil channel is defined as the second connection point. The second connection point is located downstream of the first connection point along the flow direction of the oil in the oil channel. The microfluidic system further includes a fusion electrode, which is used to apply an electric field to the second connection portion to cause the liquid in the injection channel to fuse into the droplet flowing through the second connection portion.

15. The microfluidic system for target pairing according to claim 14, characterized in that, The microfluidic system has a processing area, in which the first channel, the second channel, the oil channel, the buffer channel, the paired channel, and the injection channel are all located; The fusion electrode includes a first electrode and a second electrode, with the first electrode arranged around the processing area.

16. The microfluidic system for target pairing according to claim 15, characterized in that, The two ends of the first electrode are arranged side by side with the second electrode, and the second electrode is arranged between the two ends of the first electrode.

17. The microfluidic system for target pairing according to claim 1, characterized in that, The microfluidic system also includes an oil microvalve corresponding to the oil flow channel, which is used to control the opening and closing of the oil flow channel.

18. The microfluidic system for target pairing according to claim 1, characterized in that, The microfluidic system further includes a first microvalve control channel, the first microvalve including a diaphragm disposed between the first channel and the first microvalve control channel, the diaphragm being driven to protrude into the first channel to be in a closed state; The inner wall of the first flow channel is an arc-shaped inner wall.

19. The microfluidic system for target pairing according to claim 1, characterized in that, The microfluidic system includes a microfluidic chip, which includes a base layer, a control layer, and a flow channel layer stacked sequentially. The flow channel layer includes a first flow channel, a second flow channel, an oil flow channel, a buffer flow channel, and a paired flow channel. The control layer includes the paired microvalve, the first microvalve, the second microvalve, and the buffer microvalve.

20. The microfluidic system for target pairing according to claim 1, characterized in that, The first target is a cell, and the second target is a microsphere.

21. The microfluidic system for target pairing according to claim 20, characterized in that, The microspheres are magnetic microspheres.

22. The microfluidic system for target pairing according to claim 21, characterized in that, The microspheres are fluorescent magnetic microspheres.

23. The microfluidic system for target pairing according to claim 1, characterized in that, The first target is a first cell, and the second target is a second cell.

24. A pairing method, applied to the microfluidic system for target pairing as described in any one of claims 1 to 23, characterized in that, Includes the following steps: Identify the single first target object from the first liquid carrying the first target object, and stop the first liquid after identifying the single first target object so that the single first target object stops in the paired flow channel; Identify the single second target from the second liquid carrying the second target, and after identifying the single second target, stop the second liquid so that the single second target stops in the paired flow channel; After both the single first target and the single second target stop in the paired flow channel, the single first target and the single second target in the paired flow channel are fed into the oil through the buffer solution to form droplets.

25. The pairing method according to claim 24, characterized in that, The method for identifying a single first target object from a first liquid carrying a first target object includes the following steps: acquiring an image of a detection area, and identifying the single first target object based on image information within the image, wherein the first liquid is capable of entering the paired flow channel after passing through the detection area; And / or, the method for identifying a single second target object from a second liquid carrying a second target object includes the following steps: acquiring an image of a detection area, identifying the single second target object based on image information within the image, wherein the second liquid is capable of entering the paired flow channel after passing through the detection area.

26. The pairing method according to claim 24, characterized in that, The method for identifying a single first target object from a first liquid carrying a first target object includes the following steps: detecting a light signal within a detection area, and identifying the single first target object based on the detected light signal, wherein the first liquid is capable of entering the pairing flow channel after passing through the detection area; And / or, the method for identifying a single second target object from a second liquid carrying a second target object includes the following steps: detecting a light signal within a detection area, and identifying the single second target object based on the detected light signal, wherein the second liquid is capable of entering the paired flow channel after passing through the detection area.

27. The pairing method according to claim 24, characterized in that, The method for identifying a single first target object from a first liquid carrying a first target object includes the following steps: detecting an electrical signal generated by an electrode disposed in a detection area, and identifying the single first target object based on the detected electrical signal, wherein the first liquid is capable of entering the pairing flow channel after passing through the detection area; And / or, the method for identifying a single second target object from a second liquid carrying a second target object includes the following steps: detecting an electrical signal generated by an electrode disposed in a detection area, and identifying the single second target object based on the detected electrical signal, wherein the second liquid is capable of entering the paired flow channel after passing through the detection area.

28. The pairing method according to claim 24, characterized in that, The pairing method further includes the following steps: passing the droplet containing the single first target and the single second target through the fusion region, and applying an electric field to the fusion region to incorporate a fourth liquid into the droplet.

Images