Single cell sorting method and device

Abstract

The invention provides a single cell sorting method and device, and the method comprises the following steps: providing a thermal bubble printing head which comprises a sample adding groove, a micro-channel and a nozzle, and the sample adding groove, the micro-channel and the nozzle are sequentially communicated; applying an infiltrating reagent into the sample adding groove to infiltrate the nozzle; a cleaning reagent is applied to the sample adding groove, and the hot bubble printing head is started to print for a certain time so as to remove the infiltrating reagent remaining in the hot bubble printing head; a biological reagent is added into the sample adding groove, and the thermal bubble printing head is started to print for a certain time so as to adjust the internal environment of the thermal bubble printing head; and the cell suspension is applied to the sample adding groove, and the thermal bubble printing head is started for single cell sorting. The single cell sorting device comprises the thermal bubble printing head, the optical module and the control assembly, an automatic liquid adding system can be selectively configured for executing the single cell sorting method, and full-automatic, high-activity and continuous single cell sorting can be achieved.

Description

Technical Field

[0001] The present invention belongs to the technical fields of microfluidics, cell line development, and monoclonal antibody screening, and relates to a single cell sorting method and device. Background Art

[0002] In recent years, the biomedical industry has developed rapidly. Among them, monoclonal antibody drugs play an important role in the fields of tumors, immunity, etc. Since 2019, monoclonal antibody drugs have accounted for more than 15% of the global prescription drug share and have become the fastest-growing field in the global pharmaceutical industry. The construction of stable cell lines is the core foundation for the development of monoclonal antibody drugs, and efficient and accurate single cell sorting is the key to optimizing the process flow and increasing drug production. Due to the rapid development of the industry and the increasing production demand, it is particularly important to establish a single cell sorting system with high efficiency, high throughput, full automation, and high cell viability. Existing single cell sorting methods have problems such as low efficiency, low monoclonal rate, unstable results, and large damage to cell viability. Therefore, there is an urgent need for a single cell sorting method and device that is fully automated, has high cell viability, and high stability.

[0003] It should be noted that the above introduction of the technical background is only for the convenience of clearly and completely explaining the technical solutions of the present application and facilitating the understanding of those skilled in the art. It cannot be considered that the above technical solutions are well-known to those skilled in the art just because these solutions are described in the background art part of the present application. Summary of the Invention

[0004] In view of the above-mentioned disadvantages of the prior art, the purpose of the present invention is to provide a single cell sorting method and device, which are used to solve the problems existing in the existing single cell sorting methods, such as low efficiency, low monoclonal rate, unstable results, and large damage to cell viability.

[0005] To achieve the above object and other related objects, the present invention provides a single cell sorting method, including the following steps:

[0006] Provide a thermal inkjet printhead, the thermal inkjet printhead includes a sample addition groove, a microchannel, and a nozzle, and the sample addition groove, the microchannel, and the nozzle are sequentially connected;

[0007] Apply a wetting reagent to the sample addition groove to wet the nozzle;

[0008] Apply a cleaning reagent to the sample addition groove, and start the thermal inkjet printhead to print for a certain time to remove the residual wetting reagent inside the thermal inkjet printhead;

[0009] Apply a biological reagent to the sample addition groove, and start the thermal inkjet printhead to print for a certain time to adjust the internal environment of the thermal inkjet printhead;

[0010] Apply the cell suspension to the sample loading slot and activate the thermal bubble print head for single cell sorting.

[0011] Optionally, when the time required for single cell sorting is longer than the preset time, apply the biological reagent to the sample loading slot at specified intervals, activate the thermal bubble print head to print for a certain time to keep the inside of the thermal bubble print head clean and unobstructed, and then apply the cell suspension to the sample loading slot to continue single cell sorting.

[0012] Optionally, when the thermal bubble print head becomes blocked and / or crystallized, first apply the dirt treatment reagent to the sample loading slot and activate the thermal bubble print head to print for a certain time for dirt treatment, and then apply the cleaning reagent to the sample loading slot and activate the thermal bubble print head to print for a certain time to remove the residual dirt treatment reagent inside the thermal bubble print head. If the cleaning reagent is the same as the dirt treatment reagent, then directly apply the biological reagent to the sample loading slot subsequently without repeating the application of the cleaning reagent.

[0013] Optionally, the dirt treatment reagent includes at least one of sterile water, PBS, citric acid, and sodium hypochlorite.

[0014] Optionally, when the standby time required for the thermal bubble print head is longer than the preset time, apply the cleaning reagent to the sample loading slot and activate the thermal bubble print head to print for a certain time to keep the print head in a wetted state.

[0015] Optionally, the wetting reagent includes alcohol, the cleaning reagent includes sterile water, and the biological reagent includes PBS or cell culture medium.

[0016] Optionally, the biological reagent and the cell suspension use the same or different reagents.

[0017] Optionally, a local moisturizing module is provided on the side of the thermal bubble print head with nozzles.

[0018] Optionally, after applying the cell suspension to the sample loading slot, first use an optical module to take a photo of the side of the thermal bubble print head with nozzles to obtain a nozzle image before single cell printing, and obtain the numbers of the nozzles with single cells based on the nozzle image before single cell printing. Then, export the single cells in the corresponding nozzles to the cell receiving device in a printing manner in the preset order based on the obtained numbers of the nozzles with single cells.

[0019] Optionally, provide corresponding adapted printing reagents for different cell types to prepare cell suspensions, and provide corresponding adapted printing parameters for different printing reagents to print single cells.

[0020] Optionally, before replacing the liquid in the sample addition groove, first suck out other remaining liquids in the sample addition groove.

[0021] Optionally, applying or sucking out liquid into / from the sample addition groove is performed manually or automatically.

[0022] The present invention also provides a single-cell sorting device, including a control component, a thermal bubble print head and an optical module electrically connected to the control component. Among them, the single-cell sorting device is used to execute the single-cell sorting method described in any one of the above.

[0023] Optionally, the single-cell sorting device further includes an automatic liquid addition system electrically connected to the control component.

[0024] As described above, the single-cell sorting method of the present invention includes the following steps: providing a thermal bubble print head, which includes a sample addition groove, a microchannel and a nozzle, and the sample addition groove, the microchannel and the nozzle are sequentially connected; applying a wetting reagent to the sample addition groove to wet the nozzle; applying a cleaning reagent to the sample addition groove, and starting the thermal bubble print head to print for a certain time to remove the remaining wetting reagent inside the thermal bubble print head; applying a biological reagent to the sample addition groove, and starting the thermal bubble print head to print for a certain time to adjust the internal environment of the thermal bubble print head; applying a cell suspension to the sample addition groove, and starting the thermal bubble print head to perform single-cell sorting. The single-cell sorting device of the present invention includes a thermal bubble print head, an optical module and a control component, and optionally an automatic liquid addition system, which is used to execute the above single-cell sorting method, and can achieve fully automated, highly active and continuous single-cell sorting. Description of the Drawings

[0025] Figure 1 It shows a flowchart of the single-cell sorting method of the present invention.

[0026] Figure 2 It shows a schematic diagram when the microchannel is not wetted.

[0027] Figure 3 It shows a schematic diagram when the microchannel is wetted.

[0028] Figure 4 It shows a schematic diagram of observing the side of the thermal bubble print head with a nozzle by using an optical module.

[0029] Figure 5 It shows a specific flowchart of the single-cell sorting method of the present invention in an embodiment.

[0030] Figure 6 It shows a schematic diagram of the structure of the local humidity preservation module and its nearby structures from a first perspective in an embodiment.

[0031] Figure 7It shows a schematic structural diagram of the local moisturizing module and its nearby structures from a second perspective in one embodiment.

[0032] Explanation of reference numerals

[0033] Steps S1 to S5

[0034] 1 Thermal bubble print head

[0035] 2 Optical module

[0036] 3 Local moisturizing module Detailed implementation manners

[0037] The following uses specific examples to illustrate the implementation manners of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific implementation manners. Various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.

[0038] Please refer to Figures 1 to 7 . It should be noted that the diagrams provided in this embodiment only illustrate the basic concept of the present invention in a schematic manner. Therefore, only the components related to the present invention are shown in the diagrams, rather than being drawn according to the number, shape, and size of the components in actual implementation. The type, quantity, and ratio of each component in actual implementation can be arbitrarily changed, and the component layout type may also be more complex.

[0039] Embodiment 1

[0040] A single-cell sorting method is provided in this embodiment. Please refer to Figure 1 , which shows a flowchart of this method and includes the following steps:

[0041] S1: Provide a thermal bubble print head, the thermal bubble print head includes a sample addition groove, a microchannel, and a nozzle, and the sample addition groove, the microchannel, and the nozzle are sequentially connected;

[0042] S2: Apply a wetting reagent to the sample addition groove to wet the nozzle;

[0043] S3: Apply a cleaning reagent to the sample addition groove, and start the thermal bubble print head to print for a certain time to remove the residual wetting reagent inside the thermal bubble print head;

[0044] S4: Apply a biological reagent to the sample addition groove, and start the thermal bubble print head to print for a certain time to adjust the environment inside the thermal bubble print head;

[0045] S5: Apply a cell suspension to the sample addition groove, and start the thermal bubble print head to perform single-cell sorting.

[0046] Specifically, the thermal inkjet printhead provided in step S1 utilizes the instantaneous high temperature of the heating film in the nozzle to vaporize the liquid above the heating film, generating bubbles to push the liquid to flow and eject from the nozzle, thereby providing power for the continuous flow of the liquid. The heat generation of the heating film is controlled by the underlying circuit.

[0047] Specifically, the thermal inkjet printhead for single-cell sorting has microfluidic channels and nozzles at the micron scale. Therefore, there is a certain difficulty in wetting the channels and nozzles. The single-cell sorting method of the present invention performs a wetting treatment on the thermal inkjet printhead before printing cells to facilitate the subsequent transportation of the cell suspension and the stable generation of droplets.

[0048] As an example, in step S2, a wetting reagent with a low surface tension, such as alcohol, is dropped into the sample addition tank, and the nozzle is observed using an optical module. After confirming that all nozzles are fully wetted, the wetting reagent in the sample addition tank is aspirated.

[0049] Specifically, in the thermal inkjet printhead, the channel layer is made of a hydrophobic material. Therefore, both the surfaces of the channels and the nozzles are hydrophobic. When a wetting reagent is dropped into the sample addition tank, the liquid with a small surface tension can fill the channels at the micron scale using capillary force and enter the nozzles.

[0050] As an example, please refer to Figure 2 and Figure 3 , where Figure 2 shows a schematic diagram when the channels are not wetted, Figure 3 shows a schematic diagram when the channels are wetted.

[0051] As an example, please refer to Figure 4 , which shows a schematic diagram of observing one side of the thermal inkjet printhead 1 with nozzles using the optical module 2. In some embodiments, the optical module 2 includes a microscope, and it can be determined whether the nozzles are wetted through the image presented by the microscope. For example, in one implementation, when light irradiates the nozzles, if the image of the nozzle area is gray, it indicates that the nozzles are not wetted, and if the image of the nozzle area is white, it indicates that the nozzles are wetted.

[0052] As an example, before replacing the liquid in the sample addition tank, it is necessary to first aspirate the remaining other liquids in the sample addition tank. For example, before applying a cleaning reagent in the next step, it is necessary to first aspirate the remaining wetting reagent in the sample addition tank.

[0053] As an example, applying or sucking out liquid into / from the sample adding groove can be done manually or automatically. For example, a pipette can be manually operated to apply or suck out liquid, or a manipulator of an automatic sample adding device can be used to operate the pipette to apply or suck out liquid. Among them, if the printing method is used to drain the excess liquid in the sample adding groove, it will cause small droplets to be adsorbed on the surface of the print head to a certain extent due to excessive printing, and the state of the print head will be different from that before draining the excess liquid in the sample adding groove, which is not friendly to stable cell printing. The present invention preferably uses the method of sucking out the excess liquid in the sample adding groove with a pipette, which is beneficial to maintaining a better state of the print head.

[0054] As an example, in the step S3, sterile water is added into the sample adding groove as a cleaning reagent, and the residual wetting reagent inside the print head is removed by the spraying method of the print head.

[0055] As an example, in the step S4, phosphate buffer saline (PBS) or cell culture medium is added into the sample adding groove as a biological reagent, and the thermal bubble print head is started to print for a certain time so that the biological reagent enters the flow channel and the nozzle, which is used to adjust the environment inside the print head to be consistent with or close to the cell suspension environment.

[0056] Specifically, different biological reagents can be used according to different cell suspensions. For example, when the cells are CHO cells (Chinese hamster ovary cells), PBS, Dulbecco's Modified Eagle Medium (DMEM), CDCHO (Chinese hamster ovary cell medium), etc. can be used for cell resuspension, and serum, growth factors, etc. can be added according to needs. At this time, the biological reagent can use the same reagent as the cell suspension or not.

[0057] As an example, in the step S5, after applying the cell suspension into the sample adding groove, first use the optical module to take a picture of the side of the thermal bubble print head with the nozzle to obtain the nozzle image before single cell printing, and obtain the number of the nozzle with a single cell based on the nozzle image before single cell printing. Then, based on the obtained number of the nozzle with a single cell, the single cells in the corresponding nozzles are sequentially exported to the cell receiving device in a printing manner according to a preset order.

[0058] Specifically, according to different application requirements of single cells, there are various types of cells that need to be sorted. In the single cell sorting method of the present invention, corresponding adapted printing reagents can be provided for different cell types to prepare cell suspensions, and corresponding adapted printing parameters can be provided for different printing reagents to print single cells.

[0059] As an example, in some embodiments, the biological general reagent PBS can be selected as the printing reagent.

[0060] As an example, in some other embodiments, the cell environment reagent in the actual application scenario can be selected as the printing reagent, and the component ratio thereof can be adjusted as needed. In addition, reagents acceptable in the application scenario can be added as needed to maintain cell characteristics, so as to be more suitable for the device to perform single-cell sorting.

[0061] Specifically, the matching performance of the above reagents and the print head has formed an empirical data system through experiments, and there are adaptable printing parameters for different reagents. After multiple groups of actual project tests, this method can quickly match cell performance, so that the single cells sorted by the device have high activity.

[0062] As an example, when the time required for single-cell sorting is longer than the preset time, the biological reagent is applied to the sample addition tank at specified intervals, and the thermal bubble print head is started to print for a certain time to maintain the inside of the thermal bubble print head in a clean and smooth state, and then the cell suspension is applied to the sample addition tank to continue single-cell sorting, so as to realize continuous operation of single-cell sorting.

[0063] As an example, when the thermal bubble print head is blocked and / or crystallized, a dirt treatment reagent is first applied to the sample addition tank and the thermal bubble print head is started to print for a certain time for dirt treatment, and then the cleaning reagent is applied to the sample addition tank and the thermal bubble print head is started to print for a certain time to remove the residual dirt treatment reagent inside the thermal bubble print head. This method can effectively improve the crystallization of the print head nozzle and the dirt of the print head.

[0064] As an example, the dirt treatment reagent includes at least one of sterile water, PBS, citric acid, and sodium hypochlorite. Among them, citric acid and sodium hypochlorite can be used to lyse the residual cells in the flow channel, PBS can be used to dissolve crystals, and clean the residual cells in the flow channel, and sterile water can be used to clean the salt solution in the flow channel to clean the residual cells in the flow channel.

[0065] It should be noted that when the dirt treatment reagent uses PBS and the biological reagent also uses PBS, there is no need to apply the cleaning reagent, that is, after applying the dirt treatment reagent for dirt treatment, the cell suspension can be directly applied to the sample addition tank for single-cell sorting.

[0066] As an example, when the standby time required by the thermal bubble print head is longer than the preset time, that is, when single-cell printing is not performed for a long time, the cleaning reagent is applied to the sample addition tank, and the thermal bubble print head is started to print for a certain time to keep the print head in a wetting state, ensuring that the thermal bubble print head can be restored for use at any time.

[0067] As an example, please refer to Figure 5 , which shows the specific flowchart of the single-cell sorting method of the present invention in an embodiment. After starting, if it is determined that single-cell sorting needs to be performed on the stored content, the wetting reagent, cleaning reagent, biological reagent, and cell suspension are sequentially applied to the sample addition tank, and single-cell printing is performed. At the same time, during the single-cell printing process, it is determined whether the print head needs to be cleaned according to different situations. If necessary, the corresponding cleaning mode is selected according to the specific scenario. For example, when the time required for single-cell sorting is longer than the preset time, cleaning mode one is executed, that is, the biological reagent is applied to the sample addition tank at a specified interval time, and the thermal bubble print head is started to print for a certain time to keep the inside of the thermal bubble print head clean and unobstructed, and then the subsequent process is executed to realize the continuous operation of single-cell sorting. Another example is when the thermal bubble print head is blocked and / or crystallized, cleaning mode two is executed, that is, the dirt treatment reagent is first applied to the sample addition tank and the thermal bubble print head is started to print for a certain time for dirt treatment, then the cleaning reagent is applied to the sample addition tank and the thermal bubble print head is started to print for a certain time to remove the residual dirt treatment reagent inside the thermal bubble print head, and then the subsequent process is continued. When the standby time required by the print head is longer than the preset time, that is, when single-cell printing is not performed for a long time, the cleaning reagent is applied to the sample addition tank, and the thermal bubble print head is started to print for a certain time to keep the print head in a wetting state, ensuring that the thermal bubble print head can be restored for use at any time.

[0068] Specifically, in view of the characteristic that the biological reagent components are prone to crystallization, and the crystallization reason is related to multiple factors such as environmental temperature, environmental humidity, and air flow, the single-cell method of the present invention also provides a local humidity preservation scheme, for example Figure 4 As shown, a local humidity preservation module 3 is provided on the side of the thermal bubble print head 1 with nozzles. By locally increasing the humidity module near the print head nozzles, the humidity can be maintained constant within a certain range, thereby effectively delaying the time for the cell suspension to form crystals in the print head, and thus long-term single-cell printing operations can be maintained.

[0069] As an example, please refer to Figure 6 and Figure 7, respectively, showing the structural schematic diagrams of the local humidity preservation module 3 and its nearby structures in a first perspective and a second perspective in an embodiment. In this embodiment, the local humidity preservation module 3 includes a water tray, which is located below the thermal inkjet printhead 1 and is spaced from the thermal inkjet printhead 1 by a certain distance. The humidity near the printhead nozzles is increased by the evaporation of the water in the water tray.

[0070] As an example, a through hole is provided at the position of the water tray facing the lens of the optical module 2 to facilitate the passage of light, and the water tray can move coaxially with the lens of the optical module 2 after being installed.

[0071] As an example, the water tray can be a disposable consumable according to needs to ensure the cleanliness requirements for each use.

[0072] As an example, the water tray can also be a non-disposable component, and a sterilization method can be adopted according to needs to ensure the cleanliness requirements.

[0073] Verified by experiments, the single-cell sorting method in this embodiment will not cause biological contamination of cells, nor will it affect cell viability. It can achieve fully automatic, highly active and highly stable single-cell sorting. Among them, all nozzles of the thermal inkjet printhead are quickly wetted through a specific pretreatment process and reagents, and can maintain the wetted state for a long time; various adapted printing reagents and printing schemes are given according to different cell types, which can ensure that the cells obtained by printing have high viability; aiming at the problem of unable to continuously and efficiently sort caused by the crystallization of reagents at the nozzles, the problem of crystallization of biological reagents on the printhead can be solved by adding a local humidity preservation system, so as to support continuous printing of single cells; combined with an automatic liquid addition system, problems such as easy contamination, large error and cumbersome time-consuming in manual operation can be solved, and fully automatic continuous production can be realized.

[0074] Embodiment 2

[0075] In this embodiment, a single-cell sorting device is provided, which includes a control component and a thermal inkjet printhead and an optical module electrically connected to the control component. Among them, the single-cell sorting device is used to perform the single-cell sorting method described in Embodiment 1.

[0076] Specifically, the control component is used to control the operation of the optical module and the thermal inkjet printhead, automatically identify the nozzles with single cells and print the single cells into a single-cell receiving device, so as to achieve single-cell sorting.

[0077] As an example, the single-cell sorting device further includes an automatic liquid addition system electrically connected to the control component. For example, a liquid transfer workstation can be used to automatically realize the wetting treatment of the printhead and the sample addition operation, and realize continuous single-cell printing operations.

[0078] As an example, the single-cell sorting device further includes a computer-readable storage medium, in which programs, algorithms or instructions are stored, and are adapted to be loaded by a processor to execute the various steps of the single-cell sorting method.

[0079] Embodiment III

[0080] In this embodiment, the single-cell sorting device in Embodiment II is used for the research and development application of monoclonal antibody drugs. First, according to the cell characteristics and project requirements, a suitable printing reagent is selected to prepare a cell suspension, and the cell suspension to be sorted is adjusted to a suitable density. Install the print head in the device, and use the automatic liquid addition system to perform the print head infiltration process (apply the infiltration reagent, cleaning reagent and biological reagent in sequence). Start the device to perform single-cell sorting fully automatically. Among them, the automatic liquid addition system is responsible for the application and suction of liquids, and the optical module and the control component are responsible for the automatic recognition of single cells and printing the single cells into the well plate. This device can continuously perform single-cell sorting and can obtain results with a high monoclonal rate.

[0081] In summary, the single-cell sorting method of the present invention includes the following steps: providing a thermal bubble print head, which includes a sample addition groove, a microchannel and a nozzle, and the sample addition groove, the microchannel and the nozzle are connected in sequence; applying an infiltration reagent to the sample addition groove to infiltrate the nozzle; applying a cleaning reagent to the sample addition groove, and starting the thermal bubble print head to print for a certain time to remove the residual infiltration reagent inside the thermal bubble print head; applying a biological reagent to the sample addition groove, and starting the thermal bubble print head to print for a certain time to adjust the internal environment of the thermal bubble print head; applying a cell suspension to the sample addition groove, and starting the thermal bubble print head to perform single-cell sorting. The single-cell sorting device of the present invention includes a thermal bubble print head, an optical module and a control component, and optionally configured with an automatic liquid addition system for performing the above single-cell sorting method, and can achieve fully automatic, high-activity and continuous single-cell sorting. Therefore, the present invention effectively overcomes various disadvantages in the prior art and has high industrial utilization value.

[0082] The above embodiments are only illustrative of the principles and effects of the present invention, and are not used to limit the present invention. Any person familiar with this technology can modify or change the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or changes completed by those with ordinary knowledge in the technical field without departing from the spirit and technical ideas disclosed by the present invention should still be covered by the claims of the present invention.

Claims

1. A single-cell sorting method, characterized in that, Comprising the following steps: Providing a thermal inkjet printhead, the thermal inkjet printhead comprising a sample addition groove, a microchannel and a nozzle, the sample addition groove, the microchannel and the nozzle being sequentially connected; Applying a wetting reagent to the sample addition groove to wet the nozzle; Applying a cleaning reagent to the sample addition groove, and starting the thermal inkjet printhead to print for a certain time to remove the residual wetting reagent inside the thermal inkjet printhead; Applying a biological reagent to the sample addition groove, and starting the thermal inkjet printhead to print for a certain time to adjust the internal environment of the thermal inkjet printhead; Applying a cell suspension to the sample addition groove, and starting the thermal inkjet printhead to perform single cell sorting.

2. The single-cell sorting method according to claim 1, wherein: When the time required for single cell sorting is longer than a preset time, applying the biological reagent to the sample addition groove at specified intervals, starting the thermal inkjet printhead to print for a certain time to maintain the inside of the thermal inkjet printhead in a clean and unobstructed state, and then applying the cell suspension to the sample addition groove to continue single cell sorting.

3. The single cell sorting method according to claim 1, characterized in that When the thermal inkjet printhead is blocked and / or crystallized, first applying a dirt treatment reagent to the sample addition groove and starting the thermal inkjet printhead to print for a certain time to perform dirt treatment, and then applying the cleaning reagent to the sample addition groove and starting the thermal inkjet printhead to print for a certain time to remove the residual dirt treatment reagent inside the thermal inkjet printhead, wherein if the cleaning reagent is the same as the dirt treatment reagent, then directly applying the biological reagent to the sample addition groove subsequently without repeating the application of the cleaning reagent.

4. The single cell sorting method according to claim 3, characterized in that: The dirt treatment reagent comprises at least one of sterile water, PBS, citric acid and sodium hypochlorite.

5. The single-cell sorting method according to claim 1, wherein: When the standby time required by the thermal inkjet printhead is longer than a preset time, applying the cleaning reagent to the sample addition groove, and starting the thermal inkjet printhead to print for a certain time to keep the printhead in a wetted state.

6. The single cell sorting method according to claim 1, characterized in that: The wetting reagent comprises alcohol, the cleaning reagent comprises sterile water, and the biological reagent comprises PBS or cell culture medium.

7. The single-cell sorting method according to claim 1, characterized in that: The biological reagent and the cell suspension may use the same or different reagents.

8. The single cell sorting method according to claim 1, characterized in that: A local moisturizing module is provided on the side of the thermal inkjet printhead having the nozzle.

9. The single cell sorting method according to claim 1, characterized in that: After applying the cell suspension to the sample addition groove, first using an optical module to take a picture of the side of the thermal inkjet printhead having the nozzle to obtain a nozzle image before single cell printing, and obtaining the number of the nozzle having a single cell based on the nozzle image before single cell printing, and then sequentially exporting the single cells in the corresponding nozzles to a cell receiving device in a printing manner based on the obtained number of the nozzle having a single cell according to a preset order.

10. The single-cell sorting method according to claim 1, characterized in that: Providing corresponding adapted printing reagents for preparing cell suspensions for different cell types, and providing corresponding adapted printing parameters for printing single cells for different printing reagents.

11. The single-cell sorting method according to claim 1, wherein: Before replacing the liquid in the sample addition groove, first sucking out the remaining other liquid in the sample addition groove.

12. The single-cell sorting method according to claim 11, characterized in that: Applying or sucking out liquid into the sample addition groove is performed manually or automatically.

13. A single-cell sorting device, comprising a control component, a thermal bubble print head and an optical module electrically connected to the control component, wherein: The single cell sorting device is used to execute the single cell sorting method according to any one of claims 1-12.

14. The single-cell sorting device according to claim 13, wherein: The single cell sorting device further comprises an automatic liquid addition system electrically connected to the control component.

Images