Method for controlling tissue dissociation system, tissue dissociation system

Abstract

The embodiment of the invention relates to a method for controlling a tissue dissociation system and the tissue dissociation system. The method comprises the following steps: at a control device, controlling a tissue dissociation instrument to dissociate a tissue sample in a dissociation tube, and providing the dissociated tissue sample to a liquid workstation through a mechanical arm; treating the dissociated tissue sample by a liquid workstation, and sub-packaging the obtained cell sample into a cell counting sample plate; a mechanical arm provides the cell counting sample plate to a cell culture box, and the cell culture box performs incubation operation on cells in the cell counting sample plate; the cell counting sample plate incubated by the cell incubator is transferred to an image analysis system by the automatic guided vehicle; the image analysis system is used for carrying out cell counting on the cell counting sample plate and sending a cell counting result to the liquid workstation; and based on the cell counting result and the predetermined density value, controlling the liquid workstation to prepare the single-cell suspension with the predetermined density. In this way, tissue dissociation automation is achieved.

Description

Technical Field

[0001] This invention relates generally to the field of biological experimental control, and more specifically to methods for controlling tissue dissociation systems and tissue dissociation systems. Background Technology

[0002] In each step of preparing a single-cell suspension, it is usually necessary to manually move carriers containing tissues or cells, such as dissociation tubes, sample racks, centrifuge tubes, and sample plates, to the corresponding equipment, and manually operate equipment such as tissue dissociation instruments, liquid workstations, and image analysis systems to perform various operations on tissue samples and / or cells; the entire preparation process involves many and complex steps.

[0003] Furthermore, traditional automated biological experimental techniques only integrate the automated control of a single device, failing to address the automated experimental control of the entire single-cell suspension preparation process and multiple devices within the system. For example, existing solutions lack automated control schemes for tissue dissociation apparatuses, as well as automated control schemes for the entire tissue dissociation system in the single-cell suspension preparation process.

[0004] In summary, the shortcomings of traditional tissue dissociation methods are: many manual steps, long time consumption, and low degree of automation. Summary of the Invention

[0005] To address the aforementioned problems, this invention provides a method and a tissue dissociation system for controlling a tissue dissociation system. This system provides a highly automated tissue dissociation system, enabling the entire process from tissue dissociation to single-cell suspension preparation to be completed in a highly automated manner, thereby improving the degree of automation and experimental efficiency.

[0006] According to a first aspect of the present invention, a method for controlling a tissue dissociation system is provided. The tissue dissociation system includes a robotic arm, an unmanned transport vehicle, a dissociation tube, a sample tube rack, a tissue dissociator, a cap opener, a liquid workstation, an image analysis system, and a control device. The method includes: at the control device, controlling the tissue dissociator to dissociate a tissue sample in the dissociation tube, the dissociated tissue sample being provided to the liquid workstation via the robotic arm; processing the dissociated tissue sample via the liquid workstation to aliquot the acquired cell sample into a cell counting plate; providing the cell counting plate to a cell culture incubator via the robotic arm, so that the cell culture incubator incubates the cells in the cell counting plate based on predetermined incubation parameters; transferring the cell counting plate incubated in the cell culture incubator to the image analysis system via the unmanned transport vehicle; performing cell counting on the cell counting plate via the image analysis system to send the cell counting results to the liquid workstation; and controlling the liquid workstation to prepare a single-cell suspension of a predetermined density based on the cell counting results and a predetermined density value.

[0007] According to a second aspect of the present invention, a tissue dissociation system includes: a robotic arm configured to move a tissue sample, a dissociation tube, and / or a sample tube rack to a predetermined position in the tissue dissociation system based on control commands from a control device; a tissue dissociator configured to dissociate the tissue sample in the dissociation tube based on control commands from the control device; a liquid workstation configured to process the dissociated tissue sample to aliquot the acquired cell sample into a cell counting plate; a cell culture incubator configured to incubate cells in the cell counting plate based on predetermined incubation parameters; and an unmanned transporter. A transport vehicle is configured to transfer a cell counting sample plate incubated in a cell culture incubator to an image analysis system; the image analysis system is configured to perform cell counting on the cell counting sample plate in order to send the cell counting results to a liquid workstation; a control device is configured to control the liquid workstation to prepare a single-cell suspension of a predetermined density based on the cell counting results and a predetermined density value; and the control device is further configured to send control commands to multiple devices in a tissue dissociation system, so that each device in the system performs an operation corresponding to the control command, in order to perform the method of the first aspect of the invention.

[0008] In some embodiments, before controlling the tissue dissociation apparatus to dissociate the tissue sample in the dissociation tube, the method for controlling the tissue dissociation system further includes: transferring a sample tube holder containing the dissociation tube to a tissue dissociation temporary storage position via a robotic arm, wherein the dissociation tube contains the tissue sample to be dissociated, and the tissue dissociation temporary storage position is used to place the sample tube holder during the dissociation process; inserting the dissociation tube into the tissue dissociation apparatus at a predetermined angle so that the dissociation tube is aligned with the tube groove of the tissue dissociation apparatus, wherein the bottom of the tube groove of the sample tube holder is provided with a positioning element that aligns with the bottom of the dissociation tube; and returning the dissociation tube to the sample tube holder in response to determining that dissociation is complete. The sample holder containing the dissociated tissue sample is transferred to the capping position, which is used to stabilize the position of the sample holder when the dissociation tube is capped; the interaction position is moved along a predetermined track outside the liquid workstation, the interaction position is used to carry the dissociation tube, centrifuge tube, cell counting sample plate, cell plate, sample plate and / or sample holder, and is configured to move along the predetermined track from inside to outside the liquid workstation to a predetermined position; in response to determining that the capping machine has completed the capping operation of the dissociation tube, the sample holder containing the dissociation tube is transferred to the interaction position; and the interaction position is moved along the predetermined track into the liquid workstation.

[0009] In some embodiments, placing the dissociation tube into the tissue dissociation apparatus at a predetermined angle includes: replacing the robotic arm with a secondary gripper; removing the dissociation tube from the sample tube rack via the secondary gripper; and placing the dissociation tube into the tissue dissociation apparatus at a predetermined angle and directly aligning it with the tube slot of the dissociation apparatus.

[0010] In some embodiments, controlling the tissue dissociator to dissociate tissue samples in a dissociation tube includes: determining the operating parameters and operating time of the tissue dissociator based on predetermined dissociation parameters in order to generate dissociation control commands; and sending the generated dissociation control commands to the tissue dissociator so that the tissue dissociator runs automatically to complete the dissociation of the tissue samples.

[0011] In some embodiments, transferring a cell counting sample plate incubated in a cell culture chamber to an image analysis system via an unmanned transport vehicle includes: in response to determining that the cell counting sample incubation in the cell culture chamber is complete, transferring the incubated cell counting sample plate to a plate centrifuge via a robotic arm, so as to control the plate centrifuge to centrifuge the cell counting sample plate; and in response to the completion of centrifugation of the cell counting sample plate in the plate centrifuge, transferring the centrifuged cell counting sample plate to the unmanned transport vehicle via a robotic arm, so that the unmanned transport vehicle transfers the cell counting sample plate incubated in the cell culture chamber to the image analysis system.

[0012] In some embodiments, after the control liquid workstation prepares a single-cell suspension of a predetermined density, the method for controlling the tissue dissociation system further includes: controlling the liquid workstation to perform a mixing operation on the prepared single-cell suspension of the predetermined density; performing cell plating on the mixed single-cell suspension based on predetermined plating parameters; determining the exit position of the cell plating based on or obtaining cell plating type information and predetermined experimental attribute information; and controlling a robotic arm to transfer the cell plating to the exit position.

[0013] In some embodiments, processing the dissociated tissue sample includes: filtering the tissue sample in the dissociation tube based on predetermined operating parameters; transferring the filtered tissue sample to a centrifuge tube for multiple centrifugation, buffer addition, pipetting, and vortexing operations to obtain cell samples; and controlling the robotic arm, the interaction position, and the mechanical gripper in the liquid workstation to transfer the centrifuge tube containing the cell samples between the interaction position, predetermined positions in the liquid workstation, and the centrifuge.

[0014] In some embodiments, performing multiple centrifugation operations and adding buffer solutions on tissue samples transferred into centrifuge tubes includes: moving a sample tube rack containing a dissociation tube to a first predetermined position within the liquid workstation via a mechanical gripper to transfer the sample from the dissociation tube to a centrifuge tube containing a filter; rinsing and volume-adjusting the centrifuge tube with buffer solution; removing the filter from the volume-adjusted centrifuge tube via the mechanical gripper, and transferring the centrifuge tube with the filter removed to an interchange position; controlling the interchange position containing the centrifuge tube to move along a predetermined track to the centrifuge to transfer the centrifuge tube to a centrifuge storage position via a robotic arm; controlling the centrifuge to perform centrifugation based on predetermined centrifugation parameters in response to determining that the centrifuge tube has been transferred to the centrifuge storage position; and controlling the robotic arm to transfer the centrifuge tube after centrifugation to the interchange position so that the interchange position containing the centrifuge tube moves along a predetermined track into the liquid workstation.

[0015] In some embodiments, performing multiple centrifugation, buffer addition, pipetting, and vortexing operations on tissue samples transferred to centrifuge tubes to obtain cell samples includes: removing supernatant, adding pre-chilled cytosolic acid reagent, and performing pipetting and vortexing operations on the tissue samples in the centrifuge tubes after the current centrifugation operation; in response to determining that the room temperature incubation is complete, controlling the liquid workstation to add pre-chilled culture medium to the centrifuge tubes to terminate cytosolic acid lysis, and performing pipetting and vortexing operations; controlling the gripper in the liquid workstation to transfer the centrifuge tubes to the interaction position; controlling the interaction position to move along a predetermined track to the centrifuge, and controlling the robotic arm to transfer the centrifuge tubes to the centrifugation storage position so that the centrifuge can perform the next centrifugation operation based on predetermined centrifugation parameters.

[0016] In some embodiments, the process of performing multiple centrifugation, buffer addition, pipetting, and vortexing operations on tissue samples transferred to centrifuge tubes to obtain cell samples further includes: controlling a liquid workstation to perform supernatant removal, culture medium addition, pipetting, and vortexing operations on the centrifuge tubes after multiple centrifugation operations, and then transferring the cell samples in the centrifuge tubes to a reagent tank for cryopreservation.

[0017] In some embodiments, the tissue dissociation system further includes: an interaction position configured to carry dissociation tubes, centrifuge tubes, cell counting plates, cell plating plates, sample tube racks, and further configured to move along a predetermined track inside and outside a liquid workstation; a centrifuge configured to perform centrifugation operations on centrifuge tubes at a centrifugation storage position based on received control commands; and a control device further configured to control a robotic arm, the interaction position, and a mechanical gripper within the liquid workstation by sending control commands, such that centrifuge tubes containing cell samples are transferred between the interaction position, a predetermined position within the liquid workstation, and the centrifuge.

[0018] In some embodiments, the tissue dissociation system further includes: a sample tube rack configured to have a positioning element at the bottom of the tube slot that aligns with the bottom of the dissociation tube, so that the dissociation tube can be stored in the tube slot of the sample tube rack at a predetermined angle; and a robotic arm configured to be replaceable with a secondary gripper, which, after the dissociation tube is removed from the sample tube rack, can be placed into the tissue dissociation instrument at a predetermined angle and directly aligned with the tube slot of the dissociation instrument.

[0019] In some embodiments, the tissue dissociator is further configured to include: a communication interface configured to receive control commands from a control device and send operation return values ​​to the control device; and a dissociation controller configured to determine operating parameters and operating time of the tissue dissociator based on the received control commands, so as to automatically operate the tissue dissociator.

[0020] It should be understood that the description in this section is not intended to identify key or essential features of the embodiments of the present invention, nor is it intended to limit the scope of the invention. Other features of the invention will become readily apparent from the following description. Attached Figure Description

[0021] The above and other features, advantages, and aspects of the various embodiments of the present invention will become more apparent from the accompanying drawings and the following detailed description. In the drawings, the same or similar reference numerals denote the same or similar elements.

[0022] Figure 1 One of the schematic diagrams is shown for implementing a tissue dissociation system according to an embodiment of the present invention.

[0023] Figure 2 A second schematic diagram is shown for implementing a tissue dissociation system according to an embodiment of the present invention.

[0024] Figure 3 A third schematic diagram is shown for implementing a tissue dissociation system according to an embodiment of the present invention.

[0025] Figure 4 A fourth schematic diagram is shown for implementing a tissue dissociation system according to an embodiment of the present invention.

[0026] Figure 5 A schematic diagram of the structure of a control device according to an embodiment of the present invention is shown.

[0027] Figure 6 A flowchart of a method for controlling a tissue dissociation system according to an embodiment of the present invention is shown.

[0028] Figure 7 A flowchart of a method for transferring a dissociation tube according to an embodiment of the present invention is shown.

[0029] Figure 8A flowchart of a method for processing dissociated tissue samples according to an embodiment of the present invention is shown.

[0030] Figure 9 A block diagram of an electronic device according to an embodiment of the present invention is shown. Detailed Implementation

[0031] The following description, in conjunction with the accompanying drawings, illustrates exemplary embodiments of the present invention, including various details to aid understanding. These details should be considered merely exemplary. Therefore, those skilled in the art will recognize that various changes and modifications can be made to the embodiments described herein without departing from the scope and spirit of the invention. Similarly, for clarity and brevity, descriptions of well-known functions and structures are omitted in the following description.

[0032] The term "comprising" and its variations as used herein signify open inclusion, i.e., "including but not limited to". Unless otherwise stated, the term "or" means "and / or". The term "based on" means "at least partially based on". The terms "one example embodiment" and "one embodiment" mean "at least one example embodiment". The term "another embodiment" means "at least one additional embodiment". The terms "first", "second", etc., may refer to different or the same objects. Other explicit and implicit definitions may also be included below.

[0033] As described above, traditional automated biological experiment schemes, such as those for tissue dissociation and preparation of single-cell suspensions, only integrate the automated control of one device and cannot cover the automated experimental control of multiple experimental devices. Therefore, they cannot solve the problem of automated experimental control of multiple devices in the entire process of single-cell suspension preparation and the entire system.

[0034] In summary, the shortcomings of traditional methods for controlling tissue dissociation systems are: numerous manual steps, long processing time, and low degree of automation.

[0035] To at least partially address one or more of the aforementioned problems and other potential issues, exemplary embodiments of the present invention propose a scheme for controlling a tissue dissociation system. In this scheme, a method for controlling a tissue dissociation system and a tissue dissociation system are provided. The tissue dissociation system includes a robotic arm, a dissociation tube, a sample tube rack, a tissue dissociator, a cap opener, a liquid workstation, an image analysis system, and a control device. The control device is further configured to send control commands to multiple devices within the tissue dissociation system, causing each device in the system to perform an operation corresponding to the control command, thereby executing the method for controlling a tissue dissociation system provided in this embodiment of the invention.

[0036] The method for controlling a tissue dissociation system provided by this invention enables the control of a tissue dissociation instrument at a control device to dissociate tissue samples in a dissociation tube. The dissociated tissue samples are then provided to a liquid workstation via a robotic arm. The liquid workstation processes the dissociated tissue samples to aliquot the acquired cell samples into a cell counting plate. The robotic arm then provides the cell counting plate to a cell culture incubator, where the incubator incubates the cells based on predetermined incubation parameters. An automated guided vehicle transfers the cell counting plate incubated in the cell culture incubator to an image analysis system. The image analysis system performs cell counting on the cell counting plate and sends the cell counting results to the liquid workstation. Based on the cell counting results and a predetermined density value, the liquid workstation is controlled to prepare a single-cell suspension of a predetermined density. Therefore, this invention provides a highly automated tissue dissociation system, automating the entire process from tissue dissociation to single-cell suspension preparation, effectively improving the automation level of the tissue dissociation system, reducing manual operation, and increasing experimental efficiency.

[0037] Figure 1 A schematic diagram is shown for implementing a tissue dissociation system 100 according to an embodiment of the present invention. Figure 1 As shown, the tissue dissociation system 100 includes a robotic arm 110, a tissue dissociator 120, a cap opener 130, a liquid workstation 140, a centrifuge 150, a cell culture incubator 160, an image analysis system 170, and a control device 180.

[0038] In some embodiments, the robotic arm 110, tissue dissociation device 120, cap opener 130, liquid workstation 140, centrifuge 150, cell culture incubator 160, image analysis system 170, unmanned transport vehicle 190 and control device 180 can communicate with each other to receive control commands sent by control device 180 and to send feedback information to control device 180.

[0039] Regarding the robotic arm 110, it is configured to move tissue samples, dissociation tubes, and / or sample tube racks to predetermined positions in the tissue dissociation system based on control commands from the control device 180; the predetermined positions may include, for example, sample tube rack storage position 101, tissue dissociator 120, dissociation temporary storage position 122, cap opener 130, cap open temporary storage position 132, centrifuge 150, cell culture incubator 160, unmanned transport vehicle 190, interaction position 142, etc.

[0040] Regarding the robotic arm 110, it may, for example, move along a predetermined path (e.g., based on control commands from the control device 180 and / or based on predetermined parameters, at a predetermined time, a predetermined experimental phase, or when predetermined conditions are met) based on control commands from the control device 180. Figure 1 Move 112 in the middle to the corresponding position (e.g.) Figure 1-4 The four possible movement positions of the robotic arm are shown respectively. It should be understood that the movement positions of the robotic arm are not limited to these positions. Figure 1-4 (Illustration in the diagram) The system retrieves or places corresponding experimental items from various experimental devices within the system, ensuring that experimental items such as test tubes, sample tubes, dissociation tubes, sample plates, and sample tube racks can flow between various devices in the system 100, thereby achieving automated control of the dissociation process. It should be understood that the aforementioned experimental devices include at least the tissue dissociator 120, cap opener 130, centrifuge 150, cell culture incubator 160, liquid workstation 140, image analysis system 170, interaction station 142, unmanned transport vehicle 190, and various temporary storage locations in the system (such as 101, dissociation temporary storage station 122, cap opener 132, etc.), and should also include other experimental devices that may be added to the system 100.

[0041] Regarding the robotic arm 110, the robotic arm 110 has grippers for grasping experimental items. Figures 1-4 The robotic arm's grippers are not shown in the diagram. In order for the robotic arm to be able to grab and place the corresponding experimental items at various positions, the robotic arm can move along a predetermined track to various positions based on the control instructions of the control device 180. It can also control the lifting, rotation and movement of the robotic arm and the movement of the grippers to complete the placement and removal of experimental items in various experimental devices.

[0042] Regarding the tissue dissociation apparatus 120, it is configured to dissociate tissue samples in dissociation tubes based on control commands from the control device 180. For example, when the robotic arm 110 places a dissociation tube from the sample tube rack 200 into the tissue dissociation apparatus 120, the control device 180 sends a control command to the tissue dissociation apparatus 120, and the tissue dissociation apparatus 120 performs a dissociation operation on the sample tissue in the dissociation tube based on the received control command.

[0043] Regarding the liquid workstation 140, it is configured to process dissociated tissue samples based on control commands from the control device 180 in order to aliquot the acquired cell samples into cell counting sample plates. The liquid workstation 140 also includes an interaction position 142 and a plurality of predetermined positions (141, 142, 143, 144).

[0044] Regarding the designated locations, such as designated locations (141, 142, 143, 144), these locations are used to store various items needed for experiments, including dissociation tubes, sample tubes, sample tube racks, cell counting sample plates, sample plates, test tubes, test tube racks, centrifuge tubes, etc. Each designated location may be configured with several sub-designated locations; for example, designated location 144 includes sub-designated locations 144-1, 144-2, and 144-3. It should be understood that... Figure 1The indicated predetermined positions are merely illustrative and can be adjusted according to actual usage needs.

[0045] Regarding the interaction bit 142, it is configured to carry dissociation tubes, centrifuge tubes, cell counting sample plates, sample plates, test tubes, sample tube racks, etc., and is also configured to move along a predetermined path (e.g., based on predetermined parameters and / or control commands from control device 180). Figure 1 144) moves within and outside the liquid workstation 140, thereby enabling the interaction position 142 to move closer to the robotic arm 110, allowing the robotic arm 110 to place items into or remove items from the interaction position 142's receiving area; for example, please refer to Figure 3 The diagram shows the positions of the robotic arm 110 and the interaction position 142. At this time, the interaction position 142 has moved outside the workstation, and the robotic arm 110 can interact with the interaction position 142 to exchange items. For example, the robotic arm 110 can place the sample tube rack on the interaction position 142.

[0046] Regarding the interaction position 142, it is also configured to be movable within the liquid workstation 140, enabling a mechanical gripper (not shown in the figure) within the liquid workstation 140 to move experimental items (such as sample tube racks, dissociation tubes, sample plates, centrifuge tubes, etc.) carried on the interaction position 142 to various predetermined positions (such as 141, 142, 143, 144) within the liquid workstation 140; for example, please refer to... Figure 3 The interaction position 142 moves outside the liquid workstation 140, and the robotic arm 110 also moves to a position closer to the interaction position 142 at this time, so that the gripper of the robotic arm 110 can take away or place experimental items (such as sample tube racks) from the interaction position 142.

[0047] The cell culture incubator 160 is configured to incubate cells in a cell counting sample plate based on predetermined incubation parameters; the cell culture incubator 160 receives control commands from the control device 180 and performs incubation operations based on the control commands.

[0048] Regarding the unmanned transport vehicle 190, it is configured to transfer cell counting sample plates incubated in a cell culture incubator to an image analysis system; the unmanned transport vehicle 190 receives control commands from the control device 180 and, based on the control commands, transports the loaded experimental items (such as cell counting sample plates) to the location indicated by the control commands (such as transporting them to the image analysis system 170).

[0049] Regarding the image analysis system 170, it is configured to perform cell counting on a cell counting sample plate in order to send the cell counting results to the liquid workstation 140. For example, the image analysis system 170 uses the automated cell analysis system Celigo to perform cell counting.

[0050] Regarding the control device 180, it is configured to control the liquid workstation 140 to prepare a single-cell suspension of a predetermined density based on cell counting results and a predetermined density value; it is also configured to send control commands to a plurality of devices in the tissue dissociation system 100, so that each device in the system performs an operation corresponding to the control command, so as to perform the method for controlling the tissue dissociation system provided in the embodiments of the present invention.

[0051] The control device 180 is also configured to control the robotic arm 110, the interaction position 142, and the mechanical gripper within the liquid workstation 140 by sending control commands, so that centrifuge tubes containing cell samples are transferred between predetermined positions within the interaction position 142, the liquid workstation 140, and the centrifuge 150.

[0052] Centrifuge 150 is configured to perform centrifugation operations on centrifuge tubes in a centrifuge storage position based on received control commands; wherein, the centrifuge storage position is the location inside the centrifuge where centrifuge tubes are placed.

[0053] In some embodiments, the sample tube rack 200 is configured to have a positioning element at the bottom of the tube slot that aligns with the bottom of the dissociation tube, so that the dissociation tube can be stored in the tube slot of the sample tube rack at a predetermined angle.

[0054] In some embodiments, the robotic arm 110 is also configured to be replaceable with a secondary gripper, which allows the dissociation tube to be removed from the sample tube rack 200 and placed at a predetermined angle into the tissue dissociation apparatus 120 and directly aligned with the tube slot of the tissue dissociation apparatus 120.

[0055] In some embodiments, the tissue dissociator 120 is further configured to include: a communication interface configured to receive control commands from the control device 140 and send operation return values ​​to the control device; and a dissociation controller configured to determine operating parameters and operating time of the tissue dissociator 120 based on the received control commands, so as to automatically run the tissue dissociator to dissociate tissue samples in dissociation tubes. In some embodiments, the tissue dissociator 120 has multiple channels, enabling it to simultaneously dissociate tissue samples in multiple dissociation tubes.

[0056] The control device 180 may have one or more processing units, including dedicated processing units such as GPUs, FPGAs, and ASICs, and general-purpose processing units such as CPUs. Additionally, one or more virtual machines may run on each control device 180. In some embodiments, please refer to... Figure 5The control device 180 includes, for example, a tissue dissociation instrument control module 702, a liquid workstation control module 704, a robotic arm control module 706, an unmanned transport vehicle control module 708, a cell counting module 710, and a solution preparation module 712.

[0057] Regarding the tissue dissociator control module 702, it is used to control the tissue dissociator to dissociate the tissue sample in the dissociation tube, and the dissociated tissue sample is provided to the liquid workstation via a robotic arm.

[0058] Regarding the liquid workstation control module 704, it is used to control the liquid workstation to process the dissociated tissue sample in order to aliquot the acquired cell sample into a cell counting plate.

[0059] Regarding the robotic arm control module 706, it is used to provide the cell counting sample plate to the cell culture chamber via the robotic arm, so that the cell culture chamber can incubate the cells in the cell counting sample plate based on predetermined incubation parameters.

[0060] Regarding the unmanned transport vehicle control module 708, it is used to transfer cell counting sample plates incubated in a cell culture incubator to an image analysis system via an unmanned transport vehicle.

[0061] Regarding the cell counting module 710, it is used to count cells on a cell counting sample plate via an image analysis system so as to send the cell counting results to a liquid workstation.

[0062] Solution preparation module 712 is used to control the liquid workstation to prepare single-cell suspensions of a predetermined density based on cell count results and predetermined density values.

[0063] The following will combine Figures 6 to 8 A method for controlling a tissue dissociation system according to an embodiment of the present invention is described. Figure 6 A flowchart of a method 600 according to an embodiment of the present invention is shown. Method 600 may be performed by, for example... Figure 1 The control device 180 shown can be used to perform the operation, or it can be used in... Figure 9 The method is performed at the illustrated electronic device 900. It should be understood that method 600 may also include additional steps not shown and / or the steps shown may be omitted, and the scope of the invention is not limited in this respect.

[0064] In step 602, the control device 180 sends a control command to the tissue dissociator to control the tissue dissociator to dissociate the tissue sample in the dissociation tube. The dissociated tissue sample is then provided to the liquid workstation via a robotic arm.

[0065] In some embodiments, controlling the tissue dissociator to dissociate tissue samples in a dissociation tube includes: determining the operating parameters and operating time of the tissue dissociator based on predetermined dissociation parameters in order to generate dissociation control commands; and sending the generated dissociation control commands to the tissue dissociator so that the tissue dissociator runs automatically to complete the dissociation of the tissue samples.

[0066] In some embodiments, the tissue dissociator 120 is further configured to include: a communication interface configured to receive control commands from the control device 140 and send operation return values ​​to the control device; and a dissociation controller configured to determine operating parameters and operating time of the tissue dissociator 120 based on the received control commands, so as to automatically run the tissue dissociator to dissociate tissue samples in dissociation tubes. In some embodiments, the tissue dissociator 120 has multiple channels, enabling it to simultaneously dissociate tissue samples in multiple dissociation tubes.

[0067] Therefore, by adding a communication interface (such as an RS232 communication interface) to the tissue dissociator, the tissue dissociator can communicate with the control device 180 and receive control commands from the control device 180 to realize the automated operation of the tissue dissociator.

[0068] In step 604, the control device 180 sends a control command to the liquid workstation to process the dissociated tissue sample via the liquid workstation in order to aliquot the acquired cell sample into a cell counting plate.

[0069] Regarding the liquid workstation, it is, for example, an automated liquid workstation. The workstation has multiple predetermined locations for temporarily storing experimental equipment needed during the experiment (such as sample tubes, sample tube racks, sample plates, centrifuge tubes, etc.). The workstation also includes mechanical grippers that can move, grasp, discard, and replace grippers along a track within the workstation according to predetermined instructions and / or control instructions received from the control device 180. The automated liquid workstation, for example, includes a series of basic instructions. Users can combine these instructions according to the required experiment type and send them from the control device 180 to the liquid workstation as control commands. Alternatively, these commands may be pre-defined within the liquid workstation. For example, completing a single liquid aspiration typically requires four actions: "loading the pipette tip, aspirating, separating, and unloading the pipette tip." The liquid workstation itself has separate commands for these four actions, and the specific parameters of these commands need to be defined according to the actual experimental requirements. For example, the location, the type of pipette tip used, the type and quantity of liquid to be aspirated from which containers, the speed, whether mixing is required, whether suspension is required, the location and container to which the liquid is added, the addition height, whether mixing and shaking are required, etc. (It should be understood that the above is only an example of some commands, and the parameters included in the control commands may be more complex under actual experimental requirements.)

[0070] Therefore, the control device 180 can send control commands to the liquid workstation in advance or in real time based on the type of experiment and actual usage requirements, so that the liquid workstation can promptly process various liquids at each stage of the reaction.

[0071] In step 606, the control device 180 sends a control command to the robotic arm to provide the cell counting sample plate to the cell culture chamber via the robotic arm, so that the cell culture chamber can incubate the cells in the cell counting sample plate based on predetermined incubation parameters.

[0072] In step 608, the control device 180 sends a control command to the unmanned transport vehicle to transfer the cell counting sample plate incubated in the cell culture chamber to the image analysis system via the unmanned transport vehicle.

[0073] In some embodiments, transferring a cell counting sample plate incubated in a cell culture incubator to an image analysis system includes: in response to determining that cell counting sample incubation in the cell culture incubator is complete, transferring the incubated cell counting sample plate to a plate centrifuge via a robotic arm, so as to control the plate centrifuge to centrifuge the cell counting sample plate; and in response to the completion of centrifugation of the cell counting sample plate in the plate centrifuge, transferring the centrifuged cell counting sample plate to an unmanned transport vehicle via a robotic arm, so that the unmanned transport vehicle transfers the cell counting sample plate incubated in the cell culture incubator to the image analysis system.

[0074] Regarding the control commands sent by the control device, they can be sent to the target device based on a predetermined time to advance the experiment; or they can be sent to the target device in response to the fulfillment of predetermined conditions (for example, if the predetermined condition is that the robotic arm 110 is detected to enter a predetermined area, a control command is sent to the interaction bit 142, thereby causing the interaction bit 142 to move to one area or another area).

[0075] Regarding control commands, these may include, for example, device number, defined sample tube position, temporary storage position, and defined movement mode, action mode, execution operation, and / or program selection for the experimental equipment. In some embodiments, the movement mode of the experimental equipment may include, for example, moving along a predetermined track, moving in a certain direction, or moving along a specific trajectory; for example, moving to a predetermined position (e.g., near a temporary storage position, a position on a track, etc.). In some embodiments, the action mode of the experimental equipment, taking a robotic arm as an example, may include actions such as grasping, putting down, discarding, lifting, lowering, rotating, and corresponding amplitudes. In some embodiments, the program selection for the experimental equipment, taking an image analysis system as an example, may involve selecting Celigo for cell counting. In some embodiments, the execution operation of the experimental equipment, taking a centrifuge as an example, may include, for example, opening the hatch, closing the hatch, starting centrifugation, and sending a return value after operation. It should be understood that the above are only some examples of control commands. In the entire tissue dissociation system, there are numerous devices, and users can configure corresponding control commands for each device according to experimental needs.

[0076] Therefore, through the communication connection between the control device 180 and various experimental devices, and the design of the control commands, users can realize the automated operation of multiple experimental devices in the tissue dissociation system through control commands, thereby enabling the entire experimental process to be completed with a high degree of automation, reducing manual operation, and improving experimental efficiency and accuracy.

[0077] In step 610, the control device 180 sends a control command to the image analysis system to perform cell counting on the cell counting sample plate via the image analysis system, so as to send the cell counting results to the liquid workstation.

[0078] In step 612, the control device 180 sends a control command to the liquid workstation to control the liquid workstation to prepare a single-cell suspension of a predetermined density based on the cell count results and the predetermined density value.

[0079] In some embodiments, after the control liquid workstation prepares a single-cell suspension of a predetermined density, the method further includes: controlling the liquid workstation to perform a mixing operation on the prepared single-cell suspension of the predetermined density; performing cell plating on the mixed single-cell suspension based on predetermined plating parameters; determining the exit position of the cell plating based on or by obtaining cell plating type information and predetermined experimental attribute information; and controlling a robotic arm to transfer the cell plating to the exit position.

[0080] In summary, through the communication connection between the control device 180 and multiple experimental devices, combined with the design of control commands, users can automate the operation of multiple experimental devices in the tissue dissociation system using control commands. Furthermore, the control device 180 can determine the completion status of control commands based on the return values ​​from each experimental device, and then send control commands to the next device in the experimental process accordingly. This allows experimental materials (such as dissociation tubes, sample tube racks, centrifuge tubes, sample plates, etc.) to be transferred between multiple experimental devices in the entire tissue dissociation system via robotic arms, interactive positions, unmanned transport vehicles, and mechanical grippers in the liquid workstation, and placed into the device's operating position to complete various processing of tissue samples, cells, and liquids, thereby completing tissue dissociation and preparing the required cell suspension. Thus, the entire experimental process can be highly automated, reducing manual operation and improving experimental efficiency and accuracy.

[0081] Figure 7 A flowchart of a method 700 for transferring a dissociation tube according to an embodiment of the present invention is shown. Method 700 may be performed by, for example... Figure 1 The control device 180 shown can be used to perform the operation, or it can be used in... Figure 9 The method is performed at the illustrated electronic device 900. It should be understood that method 700 may also include additional steps not shown and / or the steps shown may be omitted, and the scope of the invention is not limited in this respect.

[0082] In step 702, the control device 180 sends a control command to the robotic arm, which then transfers the sample tube rack containing the dissociation tube to the tissue dissociation temporary storage position. The dissociation tube contains the tissue sample to be dissociated, and the tissue dissociation temporary storage position is used to place the sample tube rack during the dissociation process.

[0083] In step 704, the dissociation tube is placed into the tissue dissociation instrument at a predetermined angle via a robotic arm so that the dissociation tube is aligned with the tube slot of the tissue dissociation instrument. The bottom of the tube slot of the sample tube rack is provided with a positioning element that aligns with the bottom of the dissociation tube.

[0084] In some embodiments, placing the dissociation tube into the tissue dissociation apparatus at a predetermined angle includes: replacing the robotic arm with a secondary gripper; removing the dissociation tube from the sample tube rack via the secondary gripper; and placing the dissociation tube into the tissue dissociation apparatus at a predetermined angle and directly aligning it with the tube slot of the dissociation apparatus. Thus, the secondary gripper ensures that when the sample tube rack is placed in the dissociation storage position, the dissociation tube placed within it forms a specific angle with the tube slot of the dissociation apparatus, and that the robotic arm transfers the dissociation tube into the tube slot of the dissociation apparatus at this specific angle when transferring it to the tube slot.

[0085] Therefore, by using the positioning device at the bottom of the sample tube rack's slot, the dissociation tube can be stably placed in the sample tube rack at a specific angle. The sample tube rack containing the dissociation tube is transferred to the dissociation temporary storage position by a robotic arm. At the dissociation temporary storage position, the specific angle formed between the dissociation tube in the sample tube rack and the sample tube rack slot is the same as the specific angle when the slot in the dissociation instrument is aligned with the dissociation tube. The robotic arm only needs to remove the dissociation tube from the sample tube rack by translation and lifting, and it can be placed into the tissue dissociation instrument at a predetermined angle and directly aligned with the dissociation instrument's slot. No additional angle adjustment is required, so that the dissociation tube is directly aligned with the dissociation instrument's slot when placed into the dissociation instrument. This achieves automatic transfer of the dissociation tube and automatic operation of the dissociation instrument.

[0086] In step 706, if it is determined that dissociation is complete, the dissociation tube is placed back into the sample holder, and the sample holder containing the dissociated tissue sample is transferred to the open position, which is used to stabilize the position of the sample holder when the dissociation tube is opened.

[0087] Regarding the opening position, for example, a positioning component that matches the bottom of the sample tube rack is provided at the opening position, so that the sample tube rack can be stably positioned at a fixed angle and a fixed position at the opening position, so that the sample tube rack does not rotate or shift when the opening machine rotates the disintegration tube cap; in conjunction with the tube groove of the sample tube rack and the positioning component at the bottom of the disintegration tube, the disintegration tube body can also be stably kept in the tube rack without rotation, so that the disintegration tube cap can be opened and removed, completing the opening.

[0088] In step 708, the control device 180 sends a control command to the liquid workstation to move the interaction position along a predetermined track outside the liquid workstation. The interaction position is used to carry dissociation tubes, centrifuge tubes, cell counting sample plates, cell seeding plates, sample plates and / or sample tube racks, and is configured to move along the predetermined track from inside to outside the liquid workstation to a predetermined position.

[0089] In step 710, if the control device 180 determines that the capping machine has completed the capping operation of the dissociation tube, it sends a control command to the robotic arm, which then transfers the sample tube rack containing the dissociation tube to the interaction position.

[0090] In step 712, the control device 180 sends a control command to the liquid workstation, causing the interactive position to move along a predetermined track into the liquid workstation.

[0091] For example, please refer to Figure 2 and Figure 3 , Figure 2 This illustrates the state in which the robotic arm has moved to the vicinity of the tissue dissociation apparatus 120. Figure 3 The diagram illustrates the interaction between the robotic arm 110 and the interaction position 142. The sample tube rack 200 is located in the dissociation temporary storage position 122. After the tissue dissociation instrument 120 completes dissociation, the control device 180 sends a control command to the robotic arm 110, causing the robotic arm 110 to remove the dissociated tubes from the tissue dissociation instrument 120 and place them back into the sample tube rack 200 on the dissociation temporary storage position 122. Then, the robotic arm 110 moves the sample tube rack 200 containing the dissociated tubes to the capping position 132. The control device 180 sends a control command to the capping machine 130, causing the capping machine 130 to automatically open the caps on the sample tube rack 200. After the capping is completed, the robotic arm 110 removes the sample tube rack 200 containing the opened dissociated tubes from the opening position 142. The cover position 132 moves towards the liquid workstation. Simultaneously, the control device 180 sends a control command to the liquid workstation 140, enabling the interaction position 142 to move along a predetermined track outside the liquid workstation. This allows the robotic arm 110 to interact with the interaction position 142, which has moved outside the liquid workstation, and transfer the sample tube rack 200 onto the interaction position 142. Then, based on the control command, the interaction position 142 moves along the predetermined track to a specific position within the liquid workstation, allowing the robotic gripper within the liquid workstation to move the sample tube rack 200 and / or the dissociation tubes on it to a predetermined position within the liquid workstation for processing the dissociated tissue sample.

[0092] Therefore, by integrating a robotic arm, tissue dissociation device, cap opener, and interaction station, dissociation tubes containing tissue samples can automatically flow between the "tissue dissociation device - cap opener - interaction station - liquid workstation". The tissue dissociation is completed by the tissue dissociation device, the cap opener is completed by the cap opener, and the dissociated tissue samples are sent to the liquid workstation by the robotic arm and interaction station for subsequent processing, so that the tissue samples can be automatically dissociated, automatically opened, and automatically transferred into the liquid workstation.

[0093] Figure 8 A flowchart of a method 800 for processing dissociated tissue samples according to an embodiment of the present invention is shown. Method 800 may be performed by, for example... Figure 1 The control device 180 shown can be used to perform the operation, or it can be used in... Figure 9The method is performed at the illustrated electronic device 900. It should be understood that method 800 may also include additional steps not shown and / or the steps shown may be omitted, and the scope of the invention is not limited in this respect.

[0094] In step 802, the liquid workstation filters the tissue sample in the dissociation tube based on predetermined operating parameters.

[0095] In step 804, the liquid workstation transfers the filtered tissue sample to centrifuge tubes for multiple centrifugation, buffer addition, pipetting, and vortexing operations to obtain cell samples.

[0096] In some embodiments, some operating parameters can be pre-defined within the liquid workstation, and the control device 180 sends control commands to the liquid workstation to determine the operation to be performed and the corresponding operating parameters.

[0097] In some embodiments, performing multiple centrifugation operations and adding buffer solutions on tissue samples transferred into centrifuge tubes includes: moving a sample tube rack containing a dissociation tube to a first predetermined position within the liquid workstation via a mechanical gripper to transfer the sample from the dissociation tube to a centrifuge tube containing a filter; rinsing and volume-adjusting the centrifuge tube with buffer solution; removing the filter from the volume-adjusted centrifuge tube via the mechanical gripper, and transferring the centrifuge tube with the filter removed to an interchange position; controlling the interchange position containing the centrifuge tube to move along a predetermined track to the centrifuge to transfer the centrifuge tube to a centrifuge storage position via a robotic arm; controlling the centrifuge to perform centrifugation based on predetermined centrifugation parameters in response to determining that the centrifuge tube has been transferred to the centrifuge storage position; and controlling the robotic arm to transfer the centrifuge tube after centrifugation to the interchange position so that the interchange position containing the centrifuge tube moves along a predetermined track into the liquid workstation.

[0098] Therefore, the above solution enables automatic removal of the filter screen without manual operation; the filter screen can be removed by controlling a mechanical gripper.

[0099] In some embodiments, performing multiple centrifugation, buffer addition, pipetting, and vortexing operations on tissue samples transferred to centrifuge tubes to obtain cell samples includes: removing supernatant, adding pre-chilled cytosolic acid reagent, and performing pipetting and vortexing operations on the tissue samples in the centrifuge tubes after the current centrifugation operation; in response to determining that the room temperature incubation is complete, controlling the liquid workstation to add pre-chilled culture medium to the centrifuge tubes to terminate cytosolic acid lysis, and performing pipetting and vortexing operations; controlling the gripper in the liquid workstation to transfer the centrifuge tubes to the interaction position; controlling the interaction position to move along a predetermined track to the centrifuge, and controlling the robotic arm to transfer the centrifuge tubes to the centrifugation storage position so that the centrifuge can perform the next centrifugation operation based on predetermined centrifugation parameters.

[0100] In some embodiments, the process of performing multiple centrifugation, buffer addition, pipetting, and vortexing operations on tissue samples transferred to centrifuge tubes to obtain cell samples further includes: controlling a liquid workstation to perform supernatant removal, culture medium addition, pipetting, and vortexing operations on the centrifuge tubes after multiple centrifugation operations, and then transferring the cell samples in the centrifuge tubes to a reagent tank for cryopreservation.

[0101] In step 806, the control device 180 controls the robotic arm, the interaction position, and the mechanical gripper in the liquid workstation to transfer the centrifuge tube containing the cell sample between the interaction position, the predetermined position in the liquid workstation, and the centrifuge.

[0102] In some embodiments, the mechanical gripper is a Twister channel that can move up and down and can open and close. After gripping the sample tube, the gripper can be rotated to perform operations such as sample mixing. In addition, the filter is placed in the centrifuge tube with an outer diameter equal to that of the centrifuge tube and only a 0.5mm gap. The gripper needs to grip very precisely from the gap in order to successfully pick up and discard the cell filter.

[0103] In some embodiments, the hardware configuration of the tissue dissociation system includes a Syngren Python Junior tissue dissociator, a Hamilton Microlab StarLET liquid workstation, a custom gripper and cap opener from Huixiang, a Hettich Rotina 380 centrifuge, a Hettich Rotina 380RC centrifuge, an Aubo six-axis collaborative robotic arm, a custom Huixiang AGV mobile robot, a Perkinelmer (or Revvity) Celigo cell imager, a Thermofisher Cytomat automated cell culture incubator, a Twister Channel accessory for the Hamilton Microlab StarLET, and a custom tube rack CNC-machined according to the size and groove positions of the Syngren dissociation tubes.

[0104] Therefore, in the above method, through hardware configuration, related tube racks, grippers and other equipment structure design, combined with control command design, the tissue dissociation instrument is automatically integrated, enabling the tissue dissociation instrument to form a large automated process system together with experimental equipment such as cap openers, centrifuges, automated pipetting workstations, automated incubators, and high-precision cell imaging instruments. This achieves full automation of the tissue dissociation process and automatically prepares single-cell suspensions to prepare for subsequent implementation.

[0105] For ease of understanding, combined with Figure 1-8 A method for controlling a tissue dissociation system according to embodiments of the present invention is described.

[0106] Step 1, refer to Figure 1 Figure 2 The control device 180 sends a control command to the robotic arm 110, which then transfers the sample tube rack 200 containing the dissociation tube from the sample tube rack storage position 101 to the dissociation temporary storage position 122.

[0107] Step 2: The robotic arm 110 is replaced with a two-stage gripper, and the dissociation tube containing the tissue sample is placed into the tissue dissociation instrument 120.

[0108] Step 3: Once the dissociation tube is placed into the tissue dissociation apparatus 120, the control device 180 sends a control command to the tissue dissociation apparatus 120, and the tissue dissociation apparatus 120 starts to run automatically to dissociate the tissue.

[0109] Step 4: Once the tissue dissociation is complete, the control device 180 sends a control command to the robotic arm 110, which then places the dissociated tube back into the sample tube rack 200 on the dissociation temporary storage position.

[0110] Step 5: After the robotic arm 110 replaces the secondary gripper with the primary gripper, it transfers the sample tube rack 200 containing the dissociated tube to the opening position 132.

[0111] Step 6: Determine that the sample tube rack 200 has been transferred to the capping position 132. The control device 180 sends a control command to the capping machine 130, and the capping machine 130 completes the capping of the dissociation tube.

[0112] Step 7, refer to Figure 2 and Figure 3 Once the cap opening is confirmed, the control device 180 sends a control command to the interaction position 142, causing the interaction position 142 to move outside the liquid workstation 140. The robotic arm 110 then transfers the sample tube rack 200 containing the opened dissociation tube from the cap opening position 132 to the interaction position 142, and then transfers the sample tube rack 200 back into the liquid workstation via the interaction position 142.

[0113] Step 8: The control device 180 sends a control command to the liquid workstation 140, which uses a mechanical gripper within the liquid workstation 140 to transfer the sample tube rack 200 on the interaction position 142 to a first predetermined position (e.g., 144-1) to transfer the dissociated tissue sample from the dissociation tube to a 50ml centrifuge tube containing a filter (e.g., 5ml pipette tip treatment to prevent pipette tip clogging). Simultaneously, the filter is rinsed with 3ml of PBS buffer and the volume is adjusted (e.g., to a final volume of 8ml). Alternatively, if there is only one sample tube, the workstation performs balancing, aliquots via PBS reagent, removes the filter using the mechanical gripper, and transfers the sample tube rack containing the 50ml sample centrifuge tube to the interaction position 142. In some embodiments, the sample tube rack has at least two slots for placing sample tubes, dissociation tubes, centrifuge tubes, etc.; the tissue dissociator or centrifuge is, for example, multi-channel or dual-channel.

[0114] Step 9: The control device 180 sends a control command to the robotic arm 110, which then transfers the sample tube rack containing 50ml centrifuge tubes to the centrifugation storage position in the centrifuge 150. After confirming that the centrifuge tubes have been transferred to the centrifugation storage position, the control device 180 sends a control command to the centrifuge 150, which then performs centrifugation, for example, with centrifugation parameters of 350rcf (relative centrifugal force) and 5 minutes (open centrifugation).

[0115] Step 10: Once centrifugation is complete, the control device 180 sends a control command to the robotic arm 110, which then transfers the centrifuged 50ml centrifuge tubes, along with their sample tube racks, to the interaction position 142.

[0116] Step 11: Control device 180 sends control commands to liquid workstation 140. Liquid workstation 140 then sequentially removes the supernatant from the liquid in the centrifuged centrifuge tube, adds 4 ml of pre-chilled red lysis reagent, mixes by pipetting / twisting (gently), incubates at room temperature for 10 minutes, adds 4 ml of pre-chilled culture medium to stop red lysis, mixes by pipetting / twisting (gently), and then the centrifuge tube is transferred back to the interaction position 142 by a mechanical gripper.

[0117] Step 12: The control device 180 sends a control command to the robotic arm 110, which then transfers the sample tube rack containing 50ml centrifuge tubes to the centrifugation storage position in the centrifuge 150. After confirming that the centrifuge tubes have been transferred to the centrifugation storage position, the control device 180 sends a control command to the centrifuge 150, which then performs centrifugation, for example, with centrifugation parameters of 350rcf for 5 minutes (open centrifugation).

[0118] Step 13: Once centrifugation is complete, the control device 180 sends a control command to the robotic arm 110, which then transfers the centrifuged 50ml centrifuge tubes, along with their sample tube racks, to the interaction position 142.

[0119] Step 14: The control device 180 sends a control command to the liquid workstation 140. The liquid workstation 140 then performs the following steps on the liquid in the centrifuged centrifuge tube: remove the supernatant, add 8 ml of pre-cooled PBS buffer, mix by pipetting and twisting a few times (gently), and then rotate the centrifuge tube back to the interaction position 142 via the mechanical gripper.

[0120] Step 15: The control device 180 sends a control command to the robotic arm 110, which then transfers the sample tube rack containing 50ml centrifuge tubes to the centrifugation storage position in the centrifuge tube 150. After confirming that the centrifuge tubes have been transferred to the centrifugation storage position, the control device 180 sends a control command to the centrifuge 150, which then performs centrifugation, for example, with centrifugation parameters of 350rcf for 5 minutes (open centrifugation).

[0121] Step 16: Once centrifugation is complete, the control device 180 sends a control command to the robotic arm 110, which then connects the centrifuged 50ml centrifuge tube to its sample tube rack and transfers it to the interaction position 142.

[0122] Step 17: Control device 180 sends control commands to liquid workstation 140. Liquid workstation 140 then sequentially removes the supernatant from the liquid in the centrifuged tube, adds 2ml of culture medium, and mixes by pipetting / twist (this step requires resuspending the entire cell). The cell sample in the 50ml centrifuge tube is then transferred to the four-channel reagent tank for low-temperature preservation. Simultaneously, the cell counting chamber sample is aliquoted, and finally, the cell counting chamber sample is transferred to the interaction position 142 by a mechanical gripper.

[0123] In step 18, the control device 180 sends control commands to the robotic arm 110 and the interaction position 142 respectively, so that the interaction position 142, carrying the cell counting sample plate, moves to outside the liquid workstation, and the robotic arm 110 transfers the cell counting sample carried by the interaction position 142 to the cell culture incubator 160.

[0124] Step 19: Confirm that the cell counting sample has been transferred to the cell culture incubator 160. The control device 180 sends a control command to the cell culture incubator 160, and the cells on the cell counting sample plate are incubated in the cell culture incubator 160 (37°C for 15 min). Once the cell incubation is confirmed to be complete, the control device 180 sends a control command to the robotic arm 110, and the robotic arm 110 transfers the cell counting sample plate after cell incubation to a plate centrifuge (not shown in the figure) for centrifugation (centrifugation parameters 1 rpm, 5 min).

[0125] Step 20: After confirming that cell incubation and plate centrifugation are complete, the control device 180 sends a control command to the automated guided vehicle 190, causing the automated guided vehicle to move, thereby enabling the robotic arm 110 to transfer the cell counting sample plate to the automated guided vehicle 190 (please refer to...). Figure 4 The cell counting sample plate is transferred to the image analysis system 170 via the unmanned transport vehicle 190 for cell counting. The image analysis system 170 then sends the cell technology results to the liquid workstation 140.

[0126] Step 21: Based on the cell count results and predetermined density values, the control liquid workstation 140 prepares a single-cell suspension of predetermined density. After adjusting the suspension to the specified density and mixing it thoroughly, the cells are plated (e.g., in a 2 / 4 plate). In some embodiments, cell plate formation includes determining the type of cell plate, such as a CTG plate (CellTiter Glo Plate) or an NSG plate (Next Generation Sequencing Plate). In some embodiments, the control device 180 can also continue to send control commands to remove the cell plate and transfer it to other devices for subsequent experiments.

[0127] It is worth noting that the tissue dissociation system structure shown in the above embodiments is schematic. Figures 1-4 This is just an illustration. The positions of the various devices, robotic arms, tracks, etc. in the tissue dissociation system are also just illustrations. In actual use, the positions of the experimental devices can be adjusted according to actual needs, or other new devices can be added, as well as the movement trajectories of the robotic arms, interactive positions, and unmanned transport vehicles can be adjusted.

[0128] Figure 9 A schematic step diagram of an example electronic device 900 that can be used to implement embodiments of the contents of this specification is shown. For example, as Figure 1 The control device 180 shown can be implemented by electronic device 900. As shown, electronic device 900 includes a central processing unit (CPU) 901, which can perform various appropriate actions and processes according to computer program instructions stored in read-only memory (ROM) 902 or loaded from storage unit 908 into random access memory (RAM) 903. The random access memory 903 can also store various programs and data required for the operation of electronic device 900. The CPU 901, ROM 902, and RAM 903 are interconnected via bus 904. Input / output (I / O) interface 905 is also connected to bus 904.

[0129] Multiple components in electronic device 900 are connected to input / output interface 905, including: input unit 906, such as keyboard, mouse, microphone, etc.; output unit 907, such as various types of monitors, speakers, etc.; storage unit 908, such as disk, optical disk, etc.; and communication unit 909, such as network card, modem, wireless transceiver, etc. Communication unit 909 allows device 900 to exchange information / data with other devices through computer networks such as the Internet and / or various telecommunications networks.

[0130] The various processes and procedures described above, such as methods 600 to 800, can be executed by the central processing unit 901. For example, in some embodiments, methods 600 to 800 may be implemented as computer software programs tangibly contained in a machine-readable medium, such as storage unit 908. In some embodiments, part or all of the computer program may be loaded and / or installed on device 900 via read-only memory 902 and / or communication unit 909. When the computer program is loaded into random access memory 903 and executed by central processing unit 901, one or more of the actions of 600 to 800 described above can be performed.

[0131] This invention relates to methods, apparatus, systems, electronic devices, computer-readable storage media, and / or computer program products. The computer program product may include computer-readable program instructions for performing various aspects of the invention.

[0132] Computer-readable storage media can be tangible devices capable of holding and storing instructions for use by an instruction execution device. Computer-readable storage media can be, for example—but not limited to—electrical storage devices, magnetic storage devices, optical storage devices, electromagnetic storage devices, semiconductor storage devices, or any suitable combination thereof. More specific examples (a non-exhaustive list) of computer-readable storage media include: portable computer disks, hard disks, random access memory (RAM), read-only memory (ROM), erasable programmable read-only memory (EPROM or flash memory), static random access memory (SRAM), portable compact disc read-only memory (CD-ROM), digital multifunction disc (DVD), memory sticks, floppy disks, mechanical encoding devices, such as punch cards or recessed protrusions storing instructions thereon, and any suitable combination thereof. The computer-readable storage media used herein are not to be construed as transient signals themselves, such as radio waves or other freely propagating electromagnetic waves, electromagnetic waves propagating through waveguides or other transmission media (e.g., light pulses through fiber optic cables), or electrical signals transmitted through wires.

[0133] The computer-readable program instructions described herein can be downloaded from computer-readable storage media to various computing / processing devices, or downloaded via a network, such as the Internet, local area network, wide area network, and / or wireless network, to an external computer or external storage device. The network may include copper transmission cables, fiber optic transmission, wireless transmission, routers, firewalls, switches, gateway computers, and / or edge computing devices. A network adapter card or network interface in each computing / processing device receives the computer-readable program instructions from the network and forwards them to computer-readable storage media within the respective computing / processing device.

[0134] The computer program instructions used to perform the operations of this invention may be assembly instructions, instruction set architecture (ISA) instructions, machine instructions, machine-dependent instructions, microcode, firmware instructions, state setting data, or source code or object code written in any combination of one or more programming languages, including object-oriented programming languages ​​such as Smalltalk, C++, etc., and conventional procedural programming languages ​​such as the "C" language or similar programming languages. The computer-readable program instructions may be executed entirely on the user's computer, partially on the user's computer, as a standalone software package, partially on the user's computer and partially on a remote computer, or entirely on a remote computer or server. In cases involving a remote computer, the remote computer may be connected to the user's computer via any type of network—including a local area network (LAN) or a wide area network (WAN)—or may be connected to an external computer (e.g., via the Internet using an Internet service provider). In some embodiments, electronic circuitry, such as programmable logic circuitry, field-programmable gate arrays (FPGAs), or programmable logic arrays (PLAs), is personalized by utilizing state information from the computer-readable program instructions. This electronic circuitry can execute the computer-readable program instructions to implement various aspects of the invention.

[0135] Various aspects of the present invention are described herein with reference to flowchart illustrations and / or step diagrams of methods, apparatus (systems), and computer program products according to embodiments of the invention. It should be understood that each step in the flowchart illustrations and / or step diagrams, as well as combinations of steps in the flowchart illustrations and / or step diagrams, can be implemented by computer-readable program instructions.

[0136] These computer-readable program instructions can be provided to a processing unit of a general-purpose computer, a special-purpose computer, or other programmable data processing apparatus to produce a machine such that, when executed by the processing unit of the computer or other programmable data processing apparatus, they create means for implementing the functions / actions specified in one or more steps of the flowchart and / or diagram of steps. These computer-readable program instructions can also be stored in a computer-readable storage medium that causes a computer, programmable data processing apparatus, and / or other device to operate in a particular manner. Thus, the computer-readable medium storing the instructions comprises an article of manufacture that includes instructions for implementing aspects of the functions / actions specified in one or more steps of the flowchart and / or diagram of steps.

[0137] Computer-readable program instructions may also be loaded onto a computer, other programmable data processing apparatus, or other device to cause a series of operational steps to be performed on the computer, other programmable data processing apparatus, or other device to produce a computer-implemented process, thereby causing the instructions executed on the computer, other programmable data processing apparatus, or other device to perform the functions / actions specified in one or more steps of a flowchart and / or a diagram of steps.

[0138] The flowcharts and step diagrams in the accompanying drawings illustrate the architecture, functionality, and operation of possible implementations of systems, methods, and computer program products according to various embodiments of the present invention. In this regard, each step in the flowchart or step diagram may represent a module, segment, or part of an instruction, which contains one or more executable instructions for implementing the specified logical function. In some alternative implementations, the functions indicated in the step may occur in a different order than those indicated in the drawings. For example, two consecutive step steps may actually be executed substantially in parallel, and they may sometimes be executed in reverse order, depending on the functions involved. It should also be noted that each step in the step diagram and / or flowchart, and combinations of steps in the step diagram and / or flowchart, can be implemented using a dedicated hardware-based system that performs the specified function or action, or using a combination of dedicated hardware and computer instructions.

[0139] The various embodiments of the present invention have been described above. These descriptions are exemplary and not exhaustive, nor are they limited to the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terminology used herein is chosen to best explain the principles, practical application, or technical improvements to the embodiments in the market, or to enable others skilled in the art to understand the embodiments disclosed herein.

Claims

1. A method for controlling a tissue dissociation system, wherein the tissue dissociation system comprises a mechanical arm, a dissociation tube, a sample tube rack, a tissue dissociator, a cap opener, a liquid workstation, an image analysis system, a control device, the method comprising: controlling, at the control device, the tissue dissociator to dissociate a tissue sample in the dissociation tube, the dissociated tissue sample being provided to the liquid workstation via the mechanical arm; processing, via the liquid workstation, the dissociated tissue sample so as to aliquot the acquired cell sample to a cell counting sample plate; providing, via the mechanical arm, the cell counting sample plate to a cell incubator so as to incubate cells in the cell counting sample plate based on predetermined incubation parameters by the cell incubator; transferring, via the automated guided vehicle, the cell counting sample plate incubated by the cell incubator to the image analysis system; performing, via the image analysis system, cell counting on the cell counting sample plate so as to send cell counting results to the liquid workstation; and controlling, based on the cell counting results and predetermined density values, the liquid workstation to dispense a single cell suspension of predetermined density.

2. The method of claim 1, before controlling the tissue dissociator to dissociate the tissue sample in the dissociation tube, the method further comprising: transferring, via the mechanical arm, a sample tube rack containing the dissociation tube to a tissue dissociation staging position, the dissociation tube containing a tissue sample to be dissociated, the tissue dissociation staging position being configured to hold the sample tube rack during the dissociation process; placing the dissociation tube into the tissue dissociator at a predetermined angle so as to align the dissociation tube with a tube slot of the tissue dissociator, the tube slot of the sample tube rack being configured with a positioning member to align with a bottom of the dissociation tube; in response to determining that the dissociation is complete, placing the dissociation tube back into the sample tube rack and transferring the sample tube rack containing the dissociated tissue sample to a cap opening position, the cap opening position being configured to stabilize the sample tube rack during the cap opening process of the dissociation tube; moving an interaction position along a predetermined track outside the liquid workstation, the interaction position being configured to carry a dissociation tube, a centrifuge tube, a cell counting sample plate, a cell plating plate, a sample plate and / or a sample tube rack and move along a predetermined track to a predetermined position inside or outside the liquid workstation; in response to determining that the cap opening machine completes the cap opening process of the dissociation tube, transferring the sample tube rack containing the dissociation tube to the interaction position; and moving the interaction position along a predetermined track inside the liquid workstation.

3. The method of claim 2, wherein placing the dissociation tube into the tissue dissociator at a predetermined angle comprises: replacing the mechanical arm with a secondary gripper; and extracting the dissociation tube from the sample tube rack via the secondary gripper; and placing the dissociation tube into the tissue dissociator at a predetermined angle and directly aligning with the tube slot of the tissue dissociator.

4. The method of claim 1, wherein controlling the tissue dissociator to dissociate the tissue sample in the dissociation tube comprises: ​ ​ ​ ​ determine operation parameters and operation time of the tissue dissociator based on the predetermined dissociation parameters, so as to generate dissociation control instructions; and send the generated dissociation control instructions to the tissue dissociator, so that the tissue dissociator is automatically operated to complete dissociation of the tissue sample.

5. The method of claim 1, wherein transferring, via the automated guided vehicle, the cell counting sample plate incubated in the cell culture incubator to the image analysis system comprises: in response to determining that incubation of the cell counting sample in the cell culture incubator is completed, transferring, via the robotic arm, the cell counting sample plate after incubation to a plate centrifuge, so as to control the plate centrifuge to perform centrifugation operation on the cell counting sample plate; and in response to determining that centrifugation of the cell counting sample plate in the plate centrifuge is completed, transferring, via the robotic arm, the cell counting sample plate after centrifugation operation to the automated guided vehicle, so that the automated guided vehicle transfers the cell counting sample plate incubated in the cell culture incubator to the image analysis system.

6. The method of claim 1, after controlling the liquid handler to dispense a single cell suspension of a predetermined density, the method further comprises: controlling the liquid handler to perform mixing operation on the dispensed single cell suspension of the predetermined density; performing cell plating on the mixed single cell suspension based on predetermined plating parameters; determining a plate-out position of the cell plate based on or retrieving type information of the cell plate and predetermined experiment attribute information; controlling the robotic arm to transfer the cell plate to the plate-out position.

7. The method of claim 1, performing processing on the dissociated tissue sample comprises: performing filtration on the tissue sample in the dissociation tube based on predetermined operation parameters; transferring the filtered tissue sample to a centrifuge tube, so as to perform multiple centrifugation operations, buffer addition operations, pipetting operations, and rotation mixing operations on the tissue sample transferred to the centrifuge tube, so as to obtain a cell sample; and controlling the robotic arm, the interaction station, and a mechanical gripper in the liquid handler, so that the centrifuge tube containing the cell sample is transferred between the interaction station, a predetermined position in the liquid handler, and the centrifuge.

8. The method of claim 7, wherein performing multiple centrifugation operations, buffer addition operations on the tissue sample transferred to the centrifuge tube comprises: moving, via the mechanical gripper in the liquid handler, a sample tube rack containing the dissociation tube to a first predetermined position in the liquid handler, so as to transfer the sample in the dissociation tube to a centrifuge tube containing a filter; rinsing and setting volume of the centrifuge tube using a buffer; removing the filter in the centrifuge tube after setting volume via the mechanical gripper, and transferring the centrifuge tube after removing the filter to the interaction station; controlling the interaction station loaded with the centrifuge tube to move along a predetermined track to a centrifuge, so as to transfer the centrifuge tube to a centrifuge temporary station via a robotic arm; controlling the robotic arm to transfer the centrifuge tube to a centrifuge tube rack in the interaction station, so as to transfer the centrifuge tube to a centrifuge via a centrifuge arm. in response to determining that the centrifuge tube is transferred to the centrifuge temporary position, controlling the centrifuge to perform a centrifugation operation based on predetermined centrifugation parameters; and controlling the mechanical arm to transfer the centrifuge tube after the centrifugation is completed to the interaction position, so that the interaction position loaded with the centrifuge tube moves along a predetermined track to the liquid station.

9. The method of claim 8, wherein the multiple centrifugation operations, buffer addition operations, pipetting operations, and rotation mixing operations are performed on the tissue sample transferred to the centrifuge tube to obtain a cell sample, comprising: after performing the supernatant removal, pre-cooled lysis reagent addition, pipetting operation, and rotation mixing operation on the tissue sample in the centrifuge tube after the current centrifugation operation, performing room temperature incubation on the tissue sample in the centrifuge tube; in response to determining that the room temperature incubation is completed, controlling the liquid station to add pre-cooled culture medium to the centrifuge tube to terminate lysis, and performing pipetting operation and rotation mixing operation; controlling the gripper in the liquid station to transfer the centrifuge tube to the interaction position; controlling the interaction position to move along a predetermined track to the centrifuge, and controlling the mechanical arm to transfer the centrifuge tube to the centrifuge temporary position, so that the centrifuge is controlled to perform the next centrifugation operation based on predetermined centrifugation parameters.

10. The method of claim 9, wherein the multiple centrifugation operations, buffer addition operations, pipetting operations, and rotation mixing operations are performed on the tissue sample transferred to the centrifuge tube to obtain a cell sample, further comprising: after performing the supernatant removal, culture medium addition, pipetting operation, and rotation mixing operation on the centrifuge tube after the multiple centrifugation operations, controlling the liquid station to transfer the cell sample in the centrifuge tube to a reagent tank and perform cryopreservation.

11. A tissue dissociation system, comprising: a mechanical arm configured to move a tissue sample, a dissociation tube, and / or a sample tube rack to a predetermined position in the tissue dissociation system based on control instructions of a control device; a tissue dissociator configured to perform dissociation on a tissue sample in the dissociation tube based on control instructions of the control device; a liquid station configured to perform processing on the dissociated tissue sample to aliquot a cell sample obtained to a cell counting sample plate; a cell incubator configured to perform incubation operation on cells in the cell counting sample plate based on predetermined incubation parameters; an automated guided vehicle configured to transfer the cell counting sample plate incubated by the cell incubator to an image analysis system; the image analysis system configured to perform cell counting on the cell counting sample plate to send a cell counting result to the liquid station; the control device configured to control the liquid station to prepare a single cell suspension of a predetermined density based on the cell counting result and a predetermined density value; and the control device further configured to send control instructions to each of the devices in the tissue dissociation system respectively, so that each device in the system performs an operation corresponding to the control instruction to perform steps of the method according to any one of claims 1 to 10.

12. The system of claim 11, further comprising: ​ An interaction station configured to carry a dissociation tube, a centrifuge tube, a cell counting plate, a cell plating plate, a sample tube rack, and further configured to move along a predetermined track inside and outside the liquid station; A centrifuge configured to perform a centrifugation operation on the centrifuge tube on the centrifugation temporary station based on the received control instruction; The control device is further configured to control the mechanical arm, the interaction station, and the mechanical gripper inside the liquid station by sending the control instruction, so that the centrifuge tube containing the cell sample is transferred between the interaction station, the predetermined position inside the liquid station, and the centrifuge.

13. The system of claim 11, further comprising: A sample tube rack configured to have a positioning member at the bottom of the tube slot to align with the bottom of the dissociation tube, so that the dissociation tube can be stored in the tube slot of the sample tube rack at a predetermined angle; The mechanical arm is further configured to be replaceable with a secondary gripper, and after the dissociation tube is taken out of the sample tube rack by the secondary gripper, it can be placed in the tissue dissociation instrument at a predetermined angle and directly aligned with the tube slot of the dissociation instrument.

14. The system of claim 11, the tissue dissociation instrument is further configured to comprise: A communication interface configured to receive control instructions from the control device and send operation return values to the control device; A dissociation controller configured to determine the operation parameters and operation time of the tissue dissociation instrument based on the received control instruction, so as to automatically operate the tissue dissociation instrument.

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