Cell passage device

Abstract

The invention relates to the technical field of cell passage, and particularly discloses a cell passage device which comprises a pipetting mechanism, a material standby mechanism and an uncovering mechanism, the pipetting mechanism comprises a space mover and a pipetting assembly installed at the output end of the space mover, and the space mover is used for driving the pipetting assembly to move in the space; the material standby mechanism is used for placing test tubes, opening test tubes filled with liquid raw materials and opening standby test tubes; the cover opening machine is used for opening and closing covers of the culture dishes; the pipetting assembly comprises a pressing device, a rotating base and a plurality of pipetting guns, the rotating base and the pressing device are both installed at the output end of the space mover, the pipetting guns are distributed in the circumferential direction of the rotating base, the rotating base is used for rotating the pipetting guns to work stations, and the pressing device is used for pressing pressing heads of the pipetting guns located at the work stations to suck liquid or discharge the liquid; the pressing device is used for pressing a push head rod after the push head rod is aligned to the pipette to replace the pipette head; the pipetting mechanism further comprises a waste container located below the pipetting gun and a gun head set containing a plurality of gun heads of different specifications, the pipetting gun can be inserted into the gun heads of the gun head set to assemble new gun heads, and the gun heads or waste liquid pushed out of the pipetting gun can be received by the waste container. According to the scheme, the automatic operation of cell passage is realized, the automatic suction of the pipette and the automatic replacement of the pipette head can be realized in the pipetting process, and the control simplicity and convenience of the pipetting mechanism are ensured.

Description

Technical Field

[0001] The present invention relates to the technical field of cell passage, and in particular to a cell passage device. Background Art

[0002] Cell passaging is a key step in the cell culture process. It involves separating cells that have grown to a certain density from their original culture vessel and inoculating them proportionally into new culture vessels to maintain continued cell growth and expansion. This procedure is widely used in biomedical fields such as drug screening, gene therapy, and vaccine development.

[0003] Conventional cell passaging experiments require strict adherence to aseptic procedures. The steps typically include: using a milliliter pipette to discard the old culture medium, then using a milliliter pipette to remove PBS for washing, then using a smaller milliliter or larger microliter pipette to add a digestion solution (such as trypsin), followed by the addition of a stop solution, and then using a milliliter pipette to add complete culture medium. After centrifugation, the supernatant is removed using a milliliter pipette, and the cells are resuspended using a milliliter or microliter pipette. After resuspension, the cells are pipetted into a cell counting chamber using a small microliter pipette (e.g., 20μL), and finally, the cell suspension is aliquoted using a microliter or milliliter pipette. The entire process requires repeated changes in pipettes and corresponding tips, making it extremely cumbersome. Furthermore, due to its high professional requirements, cell passaging is typically performed by highly educated researchers in most laboratories, resulting in a significant waste of human resources.

[0004] Some high-end laboratories have designed automated equipment for cell passage. These automated equipment can replace manual operations as much as possible. For example, the "cell passage culture workstation and method" disclosed in CN116286344A realizes automated operation by integrating a pipette, a clamp and a detection probe through a robotic arm. However, this solution only has one pipette, which means that the pipette needs to meet the pumping volume requirements of all passage steps. The range span is extremely large, making it difficult to ensure accurate aspiration of different aspiration volume requirements under the passage steps; the "cell intelligent passage device" proposed in CN217868908U adopts Using modular design, the pipetting function is divided into dedicated tips (pipette tips, liquid addition tips, and waste liquid extraction tips). However, the modular design also means that when different types of pipetting are required within the same measuring range, a pipette needs to be set up for each type, resulting in a large number of pipettes required. The specific method of controlling the extraction and drainage of each pipette is not disclosed in the above-mentioned patents. In actual use, in order to ensure that the various steps of cell passage do not affect each other, the tip needs to be discarded after each use of a pipette. How to achieve the extraction and drainage of the pipette and the replacement of the pipette tip is not mentioned in the above-mentioned two patents. Summary of the Invention

[0005] The present invention aims to provide a cell passage device to realize the automated operation of cell passage and ensure that the pipetting process can realize automatic aspiration of the pipette gun and automatic replacement of the pipette gun tip, thereby ensuring the simplicity of control of the pipetting mechanism.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions: A cell passage device includes a pipetting mechanism, a material standby mechanism and a cover opening mechanism. The pipetting mechanism includes a space mover and a pipetting assembly installed at the output end of the space mover. The space mover is used to drive the pipetting assembly to move in space; the material standby mechanism is used to place test tubes, open test tubes containing liquid raw materials and open spare test tubes; the cover opening machine is used to open and close the lid of the culture dish; the pipetting assembly includes a presser, a rotating seat and multiple pipette guns. The rotating seat and the presser are all installed at the output end of the space mover. The liquid guns are distributed circumferentially on the rotating seat, which is used to rotate the pipette to the working position. The presser is used to press the pressing head of the pipette located in the working position to absorb or discharge liquid. The presser is used to press the pushing rod after aligning the pushing rod of the pipette to replace the gun tip; the pipette mechanism also includes a waste container located below the pipette and a gun tip group with multiple gun tips of different specifications. The pipette can be inserted into the gun tip of the gun tip group to assemble a new gun tip, and the gun tip or waste liquid pushed out of the pipette can be received by the waste container.

[0007] The principles and advantages of this scheme are: When adopting this scheme, when cell passaging is required, the liquid raw materials required for cell passaging, such as PBS, digestion solution, stop solution, and complete culture medium, are first placed in test tubes with tube caps, and new test tubes required for centrifugation are prepared. These test tubes are placed on the material reserve mechanism. Each time the pipetting mechanism needs to take the corresponding liquid, the culture dish is opened using the cap opener, and the caps of the test tubes are opened using the material reserve mechanism, thereby facilitating the pipette gun located in the working position of the pipetting mechanism to perform pumping and drainage actions, such as aspirating the liquid in the culture dish, aspirating the liquid in the test tube, discharging the aspirated liquid into a waste container, and adding the aspirated liquid to the culture dish / new test tube; when the pipetting mechanism is in action, since multiple pipette guns can be installed on the rotating seat of the pipetting mechanism, the choice of these pipette guns can be selected with different ranges according to the passaging needs, ensuring that passaging operations with different passaging numbers (such as 1 to 3, 1 to 5) can be met, thereby making the passaging application range wide.

[0008] Moreover, in this solution, although there are multiple pipettes in the pipette assembly, each pipette needs to rotate the rotating seat to the working position before the pipetting operation can be performed, that is, multiple pipettes share a presser, thereby simplifying the structure of the pipette assembly and also simplifying the difficulty of operating the pipette assembly.

[0009] In addition, the presser on the pipette assembly can not only press the pressing head of the pipette, thereby facilitating the extraction of the liquid by pressing, but also, with the cooperation of the rotating seat, after driving the pipette to rotate the angle, let the presser align with the push rod for replacing the pipette tip and press the push rod to realize automatic replacement of the pipette tip, meeting the need to replace a new pipette tip each time pipetting during cell passaging. When the pipette needs to use a new tip, it is only necessary to first align the used tip with the waste container, and then press the push rod so that the used tip is pushed out and falls into the waste container under the push of the push rod. Then the pipette mechanism drives the pipette to align with the required tip on the tip group and insert it downward onto the new tip, thus completing the assembly of the new tip. This solution makes the entire cell passaging device highly automated and has a wide range of applications, and the pipetting mechanism has a simple structure and low control difficulty.

[0010] Preferably, as an improvement, the presser includes a reciprocating pressure rod, a rotator and a pressing block, the reciprocating pressure rod is used to drive the pressing block away from or close to the pressing head of the pipette, the pressing block and the pressing head can fit together in a concave and convex manner, and the rotator is used to drive the pressing block to rotate, so as to act on the pressing block through the rotator, and then the pressure amount of the pipette is adjusted by screwing the pressing head with the pressing block.

[0011] Preferably, as an improvement, a plurality of suction volume data acquisition modules are provided on the rotating seat, and each suction volume data acquisition module is used to acquire the range of a corresponding pipette.

[0012] Beneficial effects: This solution facilitates the automatic adjustment of the range of different pipetting processes in different biological processing experiments through the setting of the aspiration volume data acquisition module. For example, the range of a pipette is 20-200ul. When 100ul needs to be taken, the aspiration volume of the pipette is adjusted through the rotator and the pressing block. The adjusted aspiration volume value can be directly displayed on the display of the pipette. The displayed value is collected by the aspiration volume data acquisition module. When the aspiration volume value is displayed as 100ul, the rotator is controlled by the control system to stop rotating the pressing block, thereby ensuring automatic adjustment of the aspiration volume under different usage requirements, further improving the degree of automation.

[0013] Preferably, as an improvement, a hollow cavity is provided in the middle of the rotating seat, the pipette gun is installed on the four sides of the rotating seat, and the axial ends of the hollow cavity are provided with through holes. The pipette assembly also includes a connecting rod fixedly installed on the output end of the space mover, and the free end of the connecting rod is fixed with an end plate, and a visual module is installed on the end plate. The visual module is used to collect image data below the pipette assembly.

[0014] Beneficial effects: The setting of the cavity in this scheme can not only reduce the weight of the pipetting component, but also provide a space for the wires of the electrical structure used in the pipetting component, avoiding the exposure of the wires on the periphery of the rotating seat. In addition, the suction volume data acquisition module often uses a camera. The suction volume data acquisition module can only expose a very short section on the periphery of the rotating seat or be directly installed in a hidden manner. The remaining section of the suction volume data acquisition module can be hidden in the hollow cavity, which makes the appearance of the pipetting component more concise and can also protect the suction volume data acquisition module.

[0015] In addition, in this solution, a connecting rod is set in the hollow cavity, and the end plate is fixed by the connecting rod, so that the end position corresponding to the hollow cavity can be used to install a visual module, and the image data below the pipetting component is collected by the visual module, which is equivalent to installing eyes for the pipetting component, so that after the visual module transmits the image data to the control system, it is ensured that the moving position of the pipetting component can be fully recorded, and the control system can accurately determine whether the pipetting component meets the preset requirements by judging the image data transmitted by the visual module, thereby further ensuring the accuracy of the pipetting component movement.

[0016] Preferably, as an improvement, the end plate blocks the through hole at the bottom end of the hollow cavity, the end plate can support the rotating structural part of the rotating seat, a rolling body is provided between the end plate and the bottom end surface of the hollow cavity, a plurality of rollers are installed on the end plate, the cross-section of the hollow cavity is circular, the plurality of rollers are evenly distributed along the circumference of the hollow cavity and the side surfaces of the rollers roll and rub against the inner wall of the hollow cavity.

[0017] Beneficial effect: By arranging the rolling body and the roller, the rotational stability of the rotating structural part of the rotating seat is ensured, and the end plate can also support the rotating seat, thereby improving the structural stability of the pipetting assembly.

[0018] Preferably, as an improvement, the pipette assembly further includes a plurality of pushers, each pipette gun is correspondingly mounted on the output end of a pusher, the pusher is mounted on the rotating seat, and the pusher is used to control the pipette gun to move closer to or away from the object to be sucked when the rotating seat rotates the pipette gun to the working position, so that when the spatial mover cannot accurately control the use position of the pipette gun on the pipette assembly, the pusher is used to perform more precise adjustments.

[0019] Preferably, as an improvement, it further comprises a centrifugal mechanism, and the material preparation mechanism is used to place the test tube into the centrifugal mechanism for centrifugation; The material standby mechanism includes a standby rack, a tube clamp and a tube cap opener. The standby rack can rotate and is provided with multiple test tube placement holes. Test tubes with caps can be placed on the placement holes of the standby rack and the test tubes are supported by the standby rack. The tube clamp is used to clamp the test tube body rotated to the cap opening station. The tube cap opener is used to clamp and twist open the tube cap of the test tube at the cap opening station. The space mover is used to drive the pipette gun of the pipetting assembly to extend into the test tube at the cap opening station to aspirate or inject liquid; the tube cap opener is also used to transfer the test tube between the standby rack and the centrifugal mechanism.

[0020] By setting up a material standby mechanism, the taking and placing of liquid raw materials on the standby rack can be realized automatically in conjunction with the pipetting mechanism. Each test tube is used at the capping station, and the use includes taking liquid from the test tube (such as taking PBS), injecting liquid into a new test tube (such as injecting cell fluid to be centrifuged), and taking away the test tube that needs to be centrifuged (if the test tube needs to be taken away, the tube clamper will not move, only the tube cap opener will clamp the tube cap, and the test tube to be centrifuged will be placed in the placement hole of the centrifuge mechanism under the action of the spatial manipulator attached to the tube cap opener). This solution realizes automatic centrifugation during cell passaging, ensuring that the centrifugation process can be completed automatically without human intervention.

[0021] Preferably, as an improvement, the lid opening mechanism includes a multi-axis motion actuator and a suction cup installed at the output end of the multi-axis motion actuator. The multi-axis motion actuator is used to drive the suction cup to move in space, and the spatial movement includes movement of the X-axis, Y-axis, and Z-axis. This solution enables the culture dish to be opened automatically through the setting of the lid opening mechanism, thereby further improving the degree of automation.

[0022] Preferably, as an improvement, it further includes an input and output mechanism, which is used to send the culture dish / cell counting plate into or out of the transfer operation area; the input and output mechanism includes an in-and-out material robot, an in-and-out material turntable and a receiving conveyor belt, the in-and-out material turntable includes a fixed tray and a rotatable turntable, the in-and-out material robot is used to take and place the culture dish / cell counting plate on the tray, the part of the tray facing the receiving conveyor belt is provided with a through hole, the turntable is used to switch the culture dish / cell counting plate on the tray and the receiving conveyor belt in a rotational manner, and the receiving conveyor belt is used to transport the culture dish to the bottom of the pipetting mechanism.

[0023] Beneficial effect: When this solution is adopted, when the input and output mechanism is used as the input of the culture dish or cell counting plate, the loading and unloading robot places the culture dish / cell counting plate on the loading and unloading station of the tray, and then controls the rotation of the turntable to push the culture dish / cell counting plate on the turntable on the tray until the culture dish / cell counting plate falls on the receiving conveyor belt (this position is also the transfer station of the loading and unloading turntable). After the receiving conveyor belt receives the culture dish / cell counting plate, the culture dish / cell counting plate is transported to an area convenient for the operation of the pipetting mechanism, so as to facilitate the operation of adding liquid to or extracting liquid from the culture dish with the cooperation of the pipetting mechanism and the opening mechanism. For example, if the culture dish to be subcultured is sent in, the culture dish is subjected to operations such as discarding the old culture medium, washing with PBS, adding digestion fluid, etc. For example, if it is a cell counting plate, it is moved to a position convenient for the pipette gun to be aimed at for filling, and the pipette gun can be aimed and then cell liquid is filled into the cell counting plate.

[0024] By adopting this solution, when the input and output mechanism is used as an output for culture dishes or cell counting plates, the culture dishes / cell counting plates placed on the receiving conveyor belt are transported in the reverse direction by the receiving conveyor belt to the through-hole position of the tray, and then the turntable is controlled to rotate in the reverse direction, and the culture dishes / cell counting plates are rotated to the loading and unloading station of the tray. Finally, the loading and unloading robot takes away the culture dishes / cell counting plates on the tray to realize the output of the culture dishes / cell counting plates. When outputting, the culture dishes or cell counting plates can be old culture dishes or cell counting plates or new culture dishes after the cell passage is completed.

[0025] This solution realizes the delivery or removal of culture dishes / cell counting plates to the bottom of the pipetting mechanism through the setting of the input and output mechanisms, further improving the degree of automation.

[0026] Preferably, as an improvement, it further includes a return mechanism and an integrated cell counting and cell morphology observation device. The integrated cell counting and morphology observation device can not only count cells, but also observe and record the cell morphology in the culture dish. The integrated cell counting and morphology observation device is provided with a cell image data acquisition module, which is used to collect cell image data displayed on the integrated cell counting and morphology observation device; the input and output mechanism is provided with a pusher, which is used to send the culture dish / cell counting plate on the conveyor belt to the loading platform of the integrated cell counting and morphology observation device; the return mechanism is used to return the culture dish / cell counting plate on the loading platform to the input and output mechanism.

[0027] Beneficial Effects: When this solution is adopted, the setting of the integrated cell counting and morphology observation device enables the cell morphology observation and cell counting required during the cell passaging process to be performed in the corresponding operation steps. The pusher sends the culture dish or cell counting plate filled with cell solution to the stage for real-time cell morphology observation or cell counting. After the cell morphology observation or cell counting is completed, the culture dish or cell counting plate can be returned to the receiving conveyor belt through the return mechanism. The setting of this solution further improves the degree of automation of the entire cell passaging process.

[0028] Preferably, as an improvement, the receiving conveyor belt includes synchronous, parallel, and equal-height conveyor belts, there is a spacing between the two conveyor belts, and a tilting mechanism is provided between the two conveyor belts. The tilting mechanism includes a pusher and a push rod and a baffle fixed at the output end of the pusher. The push rod is used to push the culture dish. The spacing between the push rod and the baffle is greater than the radius of the culture dish and smaller than the diameter of the culture dish. The baffle is used to tilt the culture dish against the baffle when the push rod eccentrically pushes the culture dish, so as to facilitate the pipette to absorb the liquid in the culture dish. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Schematic diagram of the three-dimensional structure of an embodiment of the present invention.

[0030] Figure 2 for Figure 1 Top view of .

[0031] Figure 3 Schematic diagram of the three-dimensional structure of the pipetting mechanism in an embodiment of the present invention.

[0032] Figure 4 for Figure 3 Schematic diagram of the three-dimensional structure of the pipette assembly when the reciprocating pressure rod moves downward to the concave-convex fit of the pressing block and the pressing head of the pipette (the spatial mover is not shown in the figure, only the pipette assembly is shown, and the pressing block and the pressing head of the pipette are matched concavely in the figure).

[0033] Figure 5 for Figure 4 Schematic diagram of the three-dimensional structure after rotation (the connecting rod and end plate are removed to facilitate the display of the hollow cavity of the rotating structure of the rotating seat).

[0034] Figure 6 for Figure 4 main view.

[0035] Figure 7 for Figure 5 Projected view toward the bottom end face of the rotating seat.

[0036] Figure 8This is a schematic diagram of the three-dimensional structure when the push rod of the pipette is facing the pressing block of the presser after the rotating base rotates a certain angle in the first embodiment of the present invention.

[0037] Figure 9 for Figure 4 A three-dimensional schematic diagram of the pipetting component after rotation at an angle, a schematic diagram of the explosion state of the connecting rod and the end plate relative to the rotating seat, and a three-dimensional combined schematic diagram of the end plate after relative rotation at an angle (the figure shows a schematic diagram of the reciprocating pressure rod constructed by the pressing drive source to rotate the gear driven by the motor, and then the reciprocating movement of the rack driven by the gear, showing that the end plate at the end of the connecting rod blocks the end of the hollow cavity, the installation position of the vision module, and the circumferential uniform distribution of the rolling elements and rollers).

[0038] Figure 10 for Figure 9 Section II in the figure.

[0039] Figure 11 for Figure 1 The three-dimensional structural diagram after the loading and unloading robot, placement rack and storage rack are not displayed.

[0040] Figure 12 for Figure 11 Schematic diagram of the three-dimensional structure without showing the pipetting mechanism.

[0041] Figure 13 This is a three-dimensional schematic diagram showing the structural relationship between the input and output mechanism, the return mechanism, and the integrated device for cell counting and cell morphology observation according to an embodiment of the present invention.

[0042] Figure 14 This is a three-dimensional structural schematic diagram showing the feed and discharge turntable, material standby mechanism, and centrifugal mechanism according to an embodiment of the present invention.

[0043] Figure 15 for Figure 14 Schematic diagram of the three-dimensional structure after rotation angle.

[0044] Figure 16 This is a three-dimensional structural diagram showing the relationship between the receiving conveyor belt, the cover opening mechanism, the pusher, the return mechanism, and the integrated device for cell counting and cell morphology observation according to an embodiment of the present invention.

[0045] Figure 17 for Figure 16 Only the top view of the second conveying section and the tilting mechanism of the receiving conveyor belt is shown.

[0046] Figure 18 Schematic diagram of the three-dimensional structure of the tilting mechanism.

[0047] Figure 19This is a front view of the process in which the tilting mechanism pushes the culture dish to an inclined position (in order to conveniently show the action process of the tilting mechanism, only the second conveying section is shown which does not block the front view of the tilting mechanism). DETAILED DESCRIPTION

[0048] The following is further described in detail through specific implementation methods: The reference numerals in the drawings of the specification include: pipetting mechanism 1, space mover 11, pipetting assembly 12, mounting base 121, Presser 122, pressing drive source 1221, reciprocating pressure rod 1222, rotator 1223, pressing block 1224, rotating seat 123, fixed structure part 1230, hollow cavity 1231, pusher 124, pipette gun 125, pressing head 1251, pushing head rod 1252, suction volume data acquisition module 126, connecting rod 127, end plate 128, visual module 129, rolling body 1281, roller 1282, gun head group 13, waste container 14, input and output mechanism 2, feeding and discharging robot 21, rotating seat 211, Z-axis linear module 212, horizontal module 213, output claw 214, placement rack 215, storage rack 216, feeding and discharging turntable 22, support Disc 221, air-avoiding groove 2211, observation hole 2212, turntable 222, air-avoiding hole 2221, receiving conveyor belt 23, pusher 24, pushing plate 241, mark recognition module 25, cover opening mechanism 3, multi-axis motion actuator 31, suction cup 32, elastic rod 33, tilting mechanism 4, pusher 41, push rod 42, push rod body 421, elastic part 422, baffle 43, material standby mechanism 5, standby rack 51, placement hole 511, tube clamp 52, tube cap opener 53, test tube 54, centrifugal mechanism 6, placement hole 61, counterweight 62, integrated cell counting and cell morphology observation equipment 7, return mechanism 8, returner 81, push-back plate 82, workbench 1000.

[0049] The embodiment is basically as shown in the attached Figures 1 to 19 shown.

[0050] Combine Figure 1 and Figure 2A cell passage device includes a pipetting mechanism 1, an input / output mechanism 2, a lid opening mechanism 3, a tilting mechanism 4, a material preparation mechanism 5, a centrifugal mechanism 6, an integrated device for cell counting and cell morphology observation 7, and a return mechanism 8. To ensure the sterility of the operation process, the pipetting mechanism 1, the input / output mechanism 2, the lid opening mechanism 3, the tilting mechanism 4, the material preparation mechanism 5, the centrifugal mechanism 6, the integrated device for cell counting and cell morphology observation 7, and the return mechanism 8 can all be installed in a sterile laboratory or in the same clean room. The clean room is equipped with a fan system, which includes a filter and is used to provide air of a set cleanliness level to the clean room. This embodiment is to illustrate the structure of the cell passage device, and the sterile laboratory or clean room is not shown in the accompanying drawings. The input / output mechanism 2, the tilting mechanism 4, the material preparation mechanism 5, the centrifugal mechanism 6, the integrated device for cell counting and cell morphology observation 7, and the return mechanism 8 are all installed on a workbench 1000.

[0051] The pipetting mechanism 1 is provided with a pipette gun 125 for performing pipetting operations during cell passage.

[0052] The input and output mechanism 2 is used to send the culture dish / cell counting plate into or out of the subculture operation area, such as into or out under the pipetting mechanism 1 to facilitate the addition of liquid, such as sending the culture dish / cell counting plate to the stage of the integrated cell morphology observation device.

[0053] The cover opening mechanism 3 is used to open or close the cover of the culture dish in the cell passage operation area.

[0054] The tilting mechanism 4 is used to push the culture dish from a horizontal position to a tilted state to facilitate the pipette 125 to suck out the liquid in the culture dish.

[0055] The material standby mechanism 5 is used to place the test tubes 54 , open the test tubes 54 containing liquid raw materials, open the standby test tubes 54 and transfer the test tubes 54 to the centrifugal mechanism 6 .

[0056] The centrifugal mechanism 6 is used to perform centrifugal operation on the cell fluid in the test tube 54 .

[0057] The integrated cell counting and cell morphology observation device 7 can be used to count cells in a cell counting chamber placed on the device's stage and observe and record the cell morphology within a culture dish. To ensure that the cell counting chamber can be accurately transferred to the stage by the input / output mechanism 2, the cell counting chamber adopts the existing structure for direct cell liquid injection, but its contour is set to a circular contour with the same periphery as the culture dish. In addition, to ensure that the pipette 125 can accurately align with the injection hole on the cell counting chamber, the counting structure of the cell counting chamber can be made larger, especially the injection hole, to facilitate alignment and injection of the pipette 125.

[0058] The return mechanism 8 is used to push the culture dish / cell counting plate on the stage of the integrated cell morphology observation device back to the input and output mechanism 2, so as to facilitate the culture dish / cell counting plate to be sent out of the cell passage operation area.

[0059] The specific structure is as follows: 1. Pipetting mechanism 1 Combine Figures 3 to 10 The pipetting mechanism 1 includes a spatial mover 11 and a pipetting assembly 12 installed at the output end of the spatial mover 11. The spatial mover 11 is used to drive the pipetting assembly 12 to move in space. The spatial mover 11 of this embodiment can be a multi-degree-of-freedom robot or a truss manipulator that can move in the X-axis, Y-axis, and Z-axis, a three-dimensional linear module, or a spatial module that moves in the X-axis and Y-axis as shown in the accompanying drawings of this embodiment. The spatial mover 11 drives the pipetting assembly 12 to move in space (such as spatial movement of the XYZ axis, such as horizontal movement of the XY axis). The spatial mover 11 only needs to meet the liquid collection and discharge requirements of the pipetting mechanism 1.

[0060] The pipetting assembly 12 includes a mounting seat 121, a presser 122, a rotating seat 123, multiple pushers 124 and multiple pipette guns 125. The mounting seat 121 is fixed to the output end of the space mover 11. The rotating seat 123 and the presser 122 are both installed on the mounting seat 121. Multiple pushers 124 are circumferentially distributed around the rotating structure of the rotating seat 123. The pressing direction of the presser 122 and the pushing direction of the pusher 124 are both parallel to the Z axis. A pipette gun 125 is installed at the output end of each pusher 124. The rotating seat 123 is used to rotate the pipette gun 125 to the working position. The pusher 124 is used to control the pipette gun 125 to approach or move away from the object to be sucked when the rotating seat 123 rotates the pipette gun 125 to the working position. The presser 122 is used to press the pipette gun 125 located in the working position to absorb or discharge liquid.

[0061] The pusher 124 is a linear module, and the output slider of the linear module can move back and forth along the Z axis. A connecting seat is fixedly installed on the output slider. The connecting seat is engaged with the pipette gun 125 and the connecting seat is provided with a strap to facilitate the disassembly and assembly of the pipette gun 125 on the connecting seat.

[0062] In addition, the multiple pipettes 125 provided with the pipette assembly 12 can be easily installed with a variety of pipettes 125 of different ranges to meet different usage requirements, such as the need to suck out the old culture medium in the culture dish during cell passaging, the need to add PBS rinse solution to the culture dish, the need to add stop solution, the need to add new complete culture medium, etc.

[0063] In addition, because the pipette 125 has a pushing rod 1252 for replacing the pipette head 125 in addition to the pressing head 1251 for aspirating liquid, every time the pipette 125 needs to replace the pipette head 125, it is only necessary to control the rotating seat 123 to rotate so that the pressing head 1251 is aligned with the pushing rod 1252, and the used pipette head 125 can be automatically pushed out, thereby improving the degree of automation of the pipette mechanism 1.

[0064] The presser 122 includes a pressing drive source 1221, a reciprocating rod 1222, a rotator 1223, and a pressing block 1224. The pressing drive source 1221 is used to drive the reciprocating rod 1222 to move along the Z axis. The rotator 1223 is fixed to the downward output end of the reciprocating rod 1222. The pressing block 1224 is fixedly mounted at the output end of the rotator 1223. The rotator 1223 is used to drive the pressing block 1224 to rotate about its axis. The reciprocating rod 1222 is used to drive the pressing block 1224 away from or toward the suction control pressing head 1251 of the pipette 125. The pressing block 1224 and the pressing head 1251 of the pipette 125 can be matched in a concave-convex manner. After the pressing block 1224 and the pressing head 1251 of the pipette 125 are matched in concave and convex manner, the rotator 1223 is started and the pressing block 1224 rotates. The pressing block 1224 realizes the screwing of the pressing head 1251 through the concave and convex matching. After the pressing head 1251 of the pipette 125 is screwed, the suction volume of the pipette 125 is adjusted, so as to facilitate the automatic adjustment of the suction volume of the pipette 125 through the rotator 1223 according to different liquid filling volume requirements, thereby improving the degree of automation. Because the pipette 125 is installed on the rotating structure of the rotating seat 123 along with the pusher 124, after the rotating seat 123 rotates to the point where the pressing block 1224 is aligned with the pushing rod 1252 for replacing the pipette tip of the pipette 125, the pressing of the pushing rod 1252 by the pressing block 1224 can push out the pipette 125 head on the pipette 125, thereby facilitating the automatic replacement of the pipette 125 head on the pipette 125.

[0065] The rotating structure of the rotating seat 123 is also provided with a suction volume data acquisition module 126 for capturing images of the suction volume of the pipette gun 125. The suction volume data acquisition module 126 uses a camera. The number of cameras in the suction volume data acquisition module 126 is the same as the number of the pipette guns 125, so that each pipette gun 125 has a corresponding camera to monitor the suction volume and ensure the accuracy of the suction volume.

[0066] A hollow cavity 1231 is provided in the middle of the rotating seat 123, and the pipette gun 125 is installed on the four sides of the rotating seat 123. The camera used by the suction volume data acquisition module 126 is installed on the side wall of the hollow cavity 1231, and the image acquisition end of the camera is hidden in the outer periphery of the rotating seat 123 or extends out of the outer periphery of the rotating seat 123 (the accompanying drawings of this embodiment take the camera extending a small section of the rotating seat 123 as an example), and the bottom of the camera is inserted into the rotating seat 123, and the connected wires and data cables are placed in the hollow cavity 1231.

[0067] The fixed structure part 1230 and the rotating structure part of the rotating seat 123 are both provided with through holes. The through hole of the rotating structure part is the above-mentioned hollow cavity 1231. Both ends of the hollow cavity 1231 have through holes. The top through hole of the hollow cavity 1231 is connected to the through hole of the fixed structure part 1230 of the rotating seat 123, so that the connecting rod 127 provided on the pipetting component 12 can pass through the entire rotating seat 123 and be fixed on the mounting seat 121. The free end of the connecting rod 127 away from the mounting seat 121 is fixed with an end plate 128, and a visual module 129 is installed on the end plate 128. The visual module 129 is used to collect image data below the pipetting component 12. The visual module 129 is close to the working position of the pipetting component 12, so that the visual module 129 can record the working process of the pipetting component 12 as much as possible. For example, the visual module 129 can collect image data of an object to be acted upon, such as a culture dish to be filled with liquid, or the position of a new gun tip that is about to be replaced, to ensure that the movement of the pipetting component 12 is more accurate. In a specific embodiment, the visual module 129 can be a camera.

[0068] In addition, the hollow cavity 1231 is a cylindrical cavity, and a plurality of evenly distributed rolling bodies 1281 are installed on the upper surface of the end plate 128 facing the bottom end surface of the rotating seat 123. The rolling bodies 1281 of this embodiment are ball bearings, and the end plate 128 provides a certain supporting force on the rotating seat 123 through the ball bearings. At the same time, the rolling of the ball bearings makes the rotation of the rotating seat 123 not affected by the end plate 128; in addition, in order to further improve the rotational stability of the rotating structure of the rotating seat 123, a plurality of rollers 1282 are installed on the end plate 128. The rollers 1282 are bearings in this embodiment. The plurality of rollers 1282 are evenly distributed along the circumference of the hollow cavity 1231 and the side surfaces of the rollers 1282 roll and rub against the inner wall of the hollow cavity 1231.

[0069] The pipetting mechanism 1 also includes a waste container 14 below the pipette 125 and a tip group 13 containing a plurality of tips of different specifications. The pipette 125 can be inserted into the tip of the tip group 13 to assemble a new tip. The tips or waste liquid pushed out of the pipette 125 can be received by the waste container 14. In order to store the waste tips and waste liquid separately, the waste container 14 is divided into two cylinders, one for receiving the waste tips and the other for receiving the waste liquid. The pipette 125 can adopt the pipette disclosed in patent publication number CN213193738U.

[0070] The automatic pipetting mechanism 1 of this embodiment is equipped with a variety of pipette guns 125 of different ranges at one time, which improves the scope of use and the demand for use. When in use, it is only necessary to control the rotating seat 123 to rotate the corresponding pipette gun 125 to the working position; and the presser 122 is used for both aspirating / discharging liquid from the pipette gun 125 in the working position and automatically pushing out the used gun tips on the pipette gun 125. A plurality of new gun tips of different specifications are also uniformly placed in the boxed gun tip group 13, which is convenient for taking gun tips of different specifications according to needs. In addition, in order to ensure the accuracy of gun tip replacement, the visual module 129 on the pipetting assembly 12 can be used to capture the position and number of the remaining gun tips in the gun tip group 13. The visual module 129 transmits the image data of the remaining gun tips to the control system, and the control system controls the spatial mover 11 to move to a position where the required gun tips can be accurately taken. Finally, the pusher 124 drives the working displacement liquid gun 125 to move downward, completing the insertion of the gun tip on the pipette gun 125.

[0071] The pipetting mechanism 1 of the entire embodiment can automatically adjust the suction volume, automatically control the suction, automatically push out the used gun tip, and automatically insert a new gun tip, which helps to achieve fully automated pipetting operations.

[0072] 2. Input / output mechanism 2, tilt mechanism 4, integrated cell counting and cell morphology observation device 7, and return mechanism 8 Combine Figures 11 to 19The input / output mechanism 2 includes an in-and-out robot 21, an in-and-out turntable 22, a receiving conveyor belt 23, and a pusher 24. The in-and-out robot 21 is a multi-degree-of-freedom robot that facilitates the transfer of culture dishes / cytometers required for cell passaging. The in-and-out robot 21 is capable of lifting culture dishes / cytometers. In this embodiment, the output claw 214 of the in-and-out robot 21 is U-shaped and capable of lifting culture dishes / round cell counting chambers. The loading and unloading robot 21 of this embodiment includes a rotating seat 211, a Z-axis linear module 212 installed at the output end of the rotating seat 211, and a horizontal module 213 installed at the output end of the Z-axis linear module 212. The output end of the horizontal module 213 is fixed with an output claw 214, so that the output hand grip can be lifted and lowered in the Z direction, rotated around the Z axis and moved in the horizontal plane to facilitate the removal and placement of culture dishes / cell counting plates from the placement rack 215 or the storage rack 216. The placement rack 215 is used to place culture dishes to be passaged, new cell counting plates, new culture dishes after passage, used cell counting plates, and used culture dishes. The storage rack 216 is used to place new culture dishes to be used. Both the placement rack 215 and the storage rack 216 can rotate to facilitate the output claw 214 to take and place culture dishes and cell counting plates in the same place.

[0073] The loading and unloading turntable 22 consists of a fixed tray 221 and a rotating turntable 222, mounted above the tray 221. The turntable 222 is equipped with a driver that controls the direction and angle of rotation of the turntable 222. The turntable 222 is equipped with at least two workstations: a loading and unloading station and a transfer station. The portion of the tray 221 corresponding to the loading and unloading station is equipped with a clearance groove 2211 for the output claw 214 of the loading and unloading robot 21 to move up and down. The portion of the tray 221 corresponding to the transfer station is equipped with a through-hole for receiving the conveyor belt 23 without clearance.

[0074] The turntable 222 is provided with an air-avoidance hole 2221 corresponding to each workstation. The air-avoidance hole 2221 can accommodate the culture dish / cell counting plate. The air-avoidance hole 2221 can also avoid the output claw 214 of the loading and unloading robot 21, so that after the air-avoidance hole 2221 on the turntable 222 is aligned with the loading and unloading workstation, the loading and unloading output claw 214 moves the supported culture dish / cell counting plate downward, and the culture dish / cell counting plate is supported by the tray 221 to realize the transfer of the culture dish / cell counting plate from the loading and unloading robot 21 to the loading and unloading workstation of the tray 221.

[0075] To improve the accuracy of cell passaging, each culture dish / cell counting plate used is marked, such as a text mark or code mark on the bottom edge of each culture dish / cell counting plate. The position of the mark should not affect the morphological observation / cell counting of cells on the culture dish / cell counting plate by the integrated cell counting and morphological observation equipment. The culture dishes / cell counting plates sent to the loading and unloading station are all marked. The tray 221 corresponding to the loading and unloading station is provided with an observation hole 2212, and a mark recognition module 25 is installed below the observation hole 2212. The mark recognition module 25 is used to identify objects placed on the tray 221 of the loading and unloading station. The mark recognition module 25 is connected to the control system to record the identity of the culture dishes / cell counting plates transferred from the loading and unloading turntable 22, thereby improving the accuracy of the control. At the same time, it is convenient to know the passaging status of the corresponding culture dish in a marked manner after the cell passaging is completed.

[0076] The tag recognition module 25 in this embodiment can be a camera or a radio frequency reader connected to the control system.

[0077] The receiving conveyor belt 23 includes a first conveying section and a second conveying section that are parallel and arranged along the conveying direction. The first conveying section is a single conveyor belt located at the transfer station, and the second conveying section is immediately connected to the end of the first conveying section. The second conveying section includes two parallel, equal-height and synchronized conveyor belts. The receiving conveyor belt 23 is used to support and convey culture dishes / cell counting plates.

[0078] The tilting mechanism 4 is arranged between the two conveyor belts of the second conveyor section. Figures 16 to 19The tilting mechanism 4 includes a pusher 41 and a push rod 42 and a baffle 43 fixed at the output end of the pusher 41. The pusher 42 and the baffle 43 are located between the two conveyor belts of the first conveying section. The pusher 41 drives the pusher 42 and the baffle 43 to rise and fall synchronously along the Z axis. The pusher 42 is used to push the culture dish. The distance between the pusher 42 and the baffle 43 is greater than the radius of the culture dish and smaller than the diameter of the culture dish. The baffle 43 is used to tilt the culture dish against the baffle 43 when the pusher 42 eccentrically pushes the culture dish, so as to facilitate the pipette 125 to absorb the liquid in the culture dish. In this embodiment, the baffle 43 is an arc-shaped plate. Cooperating with the arc structure of the culture dish, it further ensures that after the push rod 42 pushes the culture dish to tilt, the culture dish will not fall in other directions. The push rod 42 is an elastic push rod 42, which includes a push rod body 421 and an elastic member 422. The push rod body 421 is slidably connected to the output end of the pusher 41 along the Z axis, and the elastic member 422 is arranged between the push rod body 421 and the output end of the pusher 41. The elastic member 422 of this embodiment is a spring, which is sleeved on the push rod body 421. One end of the elastic member 422 abuts against the push rod body 421, and the other end abuts against the output end of the pusher 41. The provision of the elastic push rod 42 allows the push rod 42 to slowly push the culture dish using its elasticity, avoiding overturning the culture dish due to excessive pushing.

[0079] The pusher 24 of the input and output mechanism 2 is installed above the end of the second conveying section away from the first conveying section. The pusher 24 is located below the pipetting mechanism 1. The pusher 24 is used to deliver the culture dish / cell counting plate of the second conveying section to the stage of the integrated cell counting and morphological observation device.

[0080] The pusher 24 includes a pushing conveyor belt and a pushing plate 241 fixed on the pushing conveyor belt. A transition support platform is provided between the integrated cell counting and cell morphology observation device 7 and the receiving conveyor belt 23. The height of the transition support platform is the same as that of the receiving conveyor belt 23. After the pushing conveyor belt is started, the pushing plate 241 is used to push the culture dish / cell counting plate on the second conveying section to the loading platform of the integrated cell counting and cell morphology observation device 7 through the transition support platform. After the culture dish / cell counting plate is placed on the loading platform, the integrated cell counting and cell morphology observation device 7 can conveniently perform cell morphology observation or cell counting.

[0081] The return mechanism 8 includes a fixedly installed return device 81 and a push-back plate 82 installed at the output end of the return device 81. The return device 81 is used to drive the push-back plate 82 away from or close to the pusher 24. In this embodiment, the return device 81 adopts a linear motor or a cylinder. The return mechanism 8 is used to push the culture dish / cell counting plate on the stage back to the input and output mechanism 2.

[0082] Through the coordinated design of the pusher 24 and the return mechanism 8, the culture dish / cell counting plate can be transferred between the input and output mechanism 2 and the integrated cell counting and cell morphology observation device 7, which facilitates the transfer of the culture dish / cell counting plate again after automatic morphological observation or cell counting.

[0083] The integrated cell counting and morphological observation device can not only count cells, but also observe and record the cell morphology in the culture dish. The integrated cell counting and morphological observation device is provided with a cell image data acquisition module, which is used to acquire cell image data displayed on the integrated cell counting and morphological observation device; the cell image data acquisition module is connected to a control system, and the cell image data acquisition module is used to acquire clear cell image data obtained after the integrated cell counting and morphological observation device automatically adjusts the focus in real time. The cell image data acquisition module is used to transmit the acquired real-time cell image data to the control system. The control system includes an analysis mode selection, which includes morphological observation and cell counting. The control system is used to count the total number of cells in the cell counting plate after the analysis mode is selected as cell counting, and the control system is used to calculate the cell rounding rate of the cells in the culture dish after the analysis mode is selected as morphological observation.

[0084] The control system is used to analyze the cell rounding rate in the real-time cell image data at equal intervals or in real time, and display and store the cell rounding rate over time on a display screen connected to the control system (display / store a curve of the cell rounding rate over time). The image data acquisition module can use a CCD camera.

[0085] 3. Opening mechanism 3 Combine Figure 16 The lid opening mechanism 3 includes a multi-axis motion actuator 31 and a suction cup 32 installed at the output end of the multi-axis motion actuator 31. The multi-axis motion actuator 31 is fixedly installed and is used to drive the suction cup 32 to move along the X-axis, Y-axis, and Z-axis. The multi-axis motion actuator 31 can specifically adopt a three-axis linear module. The suction cup 32 is connected to the negative pressure so that the suction cup 32 can open the lid of the culture dish through the multi-axis motion actuator 31 after being adsorbed to the lid of the culture dish, making it convenient for the pipetting mechanism 1 to add or absorb liquid to the culture dish.

[0086] In this embodiment, in order to facilitate the passaging of cells after cell morphology observation, the number of suction cups 32 is set to multiple, and the negative pressure of each suction cup 32 is connected without affecting each other, so that each suction cup 32 can absorb the lid of the culture dish at the corresponding position, making it convenient for the opening mechanism 3 to open the lids of multiple culture dishes at one time.

[0087] In addition, because the lid opening mechanism 3 can move in the XYZ axis, when the cell passaging is completed and the culture dish needs to be evenly filled with cell fluid, the suction cup 32 on the lid opening mechanism 3 can be used to firmly hold the lid of the corresponding culture dish, and then the multi-axis motion actuator 31 can be controlled to drive the suction cup 32 to move in the horizontal plane to simulate the manual cross movement of the culture dish, thereby achieving uniform spreading of the cell fluid in the culture dish.

[0088] In addition, an elastic rod 33 is provided between the suction cup 32 and the output end of the multi-axis motion actuator 31 of this embodiment. The deformation direction of the elastic rod 33 is the Z axis. The elastic rod 33 includes a sliding rod slidably connected to the output end of the multi-axis motion actuator 31. A spring is sleeved on the sliding rod. One end of the spring is against the output end of the actuator, and the other end is against the sliding rod. The free end of the sliding rod is fixed to the suction cup 32. This solution allows the elastic rod 33 to buffer the process of the multi-axis motion actuator 31 driving the suction cup 32 to press down on the lid of the culture dish, thereby avoiding the problem of the multi-axis motion actuator 31 pressing down a slightly larger distance and crushing or damaging the culture dish.

[0089] 4. Material standby mechanism 5 and centrifugal mechanism 6 Combine Figure 14 and Figure 15 The material standby mechanism 5 includes a standby rack 51, a tube clamp 52 and a tube cap opener 53. The standby rack 51 can rotate. The rotation of the standby rack 51 is controlled by a rotary driver. The standby rack 51 is in the shape of a truncated cone. A plurality of test tube 54 placement holes 511 are provided on the circumferential edge of the truncated cone of the standby rack 51. The test tubes 54 with tube caps can be placed on the placement holes 511 of the standby rack 51 and the test tubes 54 are supported by the standby rack 51. Because the size of the test tube 54 cap is larger than the test tube body 54, when the test tube cap 54 is not opened, the test tube 54 is equivalent to hanging on the placement hole 511.

[0090] The tube gripper 52 is used to grip the tube body of the test tube 54 rotated to the uncapping station. The tube gripper 52 of this embodiment can adopt a finger cylinder or an electric gripper. The tube gripper 52 includes two gripping fingers that can move closer to or away from each other. The two gripping fingers move closer to each other to clamp the test tube 54. After the two gripping fingers move away from each other, the test tube 54 can shuttle between the two gripping fingers as the spare rack 51 rotates.

[0091] The tube cap opener 53 is used to open the tube cap of the test tube 54 at the cap opening station. The tube cap opener 53 includes a space manipulator and a tube cap opening structure installed at the output end of the space manipulator. The space manipulator is used to drive the tube cap opening structure to move in three-dimensional space. The tube cap opening structure is used to screw the tube cap of the test tube 54. The specific structure can refer to the tube cap opener 53 composed of a multi-degree-of-freedom robot and a drive-control integrated opening device in the intelligent opening robot for new coronavirus sampling tubes disclosed in CN113003506A.

[0092] The spatial mover 11 is used to drive the pipette gun 125 of the pipetting assembly 12 to extend into the test tube 54 at the capping station to suck or inject liquid.

[0093] The centrifuge mechanism 6 includes a rotatable centrifuge frame with a receiving hole 61 and a counterweight 62 symmetrically arranged about the centrifuge frame's rotation axis. The counterweight 62 ensures stable rotation during centrifugation, even when only a single test tube 54 is being centrifuged, thereby ensuring the centrifugal effect. A cap opener 53 is used to place the test tube 54 to be centrifuged from the standby rack 51 into the receiving hole 61. After centrifugation is complete, the cap opener 53 is used to return the centrifuged test tube 54 to the standby rack 51. The cap of the centrifuged test tube 54 is then removed by the cooperation of the tube gripper 52 and the cap opener 53.

[0094] This embodiment also provides a cell passaging method, comprising the following steps: S1. The culture dish to be subcultured is sent to the inlet and outlet station of the inlet and outlet turntable 22 through the input and output mechanism 2; the turntable 222 on the inlet and outlet turntable 22 rotates to rotate the culture dish to be subcultured to the transfer station, and the culture dish automatically falls onto the receiving conveyor belt 23.

[0095] S2. The culture dish to be subcultured is transported by the receiving conveyor belt 23 toward the integrated device for cell counting and cell morphology observation 7, and the pusher 24 pushes the culture dish to be subcultured onto the integrated device for cell counting and cell morphology observation 7 for cell morphology observation. The control system displays and records the cell production morphology of the culture dish.

[0096] S3. The culture dish on the integrated cell counting and cell morphology observation device 7 is returned to the input / output mechanism 2 by the return mechanism 8 , and the input / output mechanism 2 drives the culture dish to move below the pipetting mechanism 1 .

[0097] S4. Use the cover opening mechanism 3 to open the cover of the culture dish to be subcultured.

[0098] S5: Use the pipetting mechanism 1 to perform the following operations on the culture dish to be subcultured: S51, remove the old culture medium from the culture dish; S52, injecting PBS buffer into the culture dish to wash the cells; S53, remove the PBS buffer and add digestion solution; In S51 and S52, in order to extract the liquid in the culture dish as much as possible (such as old culture medium or PBS buffer), before the tip of the pipette 125 is inserted into the culture dish, the tilting mechanism 4 is controlled to push the culture dish to a tilted state to facilitate the removal of liquid.

[0099] S6. Control the lid opening mechanism 3 to close the lid of the culture dish, and then control the input and output mechanism 2 to send the culture dish to the integrated cell counting and cell morphology observation device 7 for morphological observation. During this process, the cell image data acquisition module collects clear cell image data; during the cell morphology observation process, the pipetting mechanism 1 adjusts the required pipette gun 125 and uses a new pipette tip to absorb the required stop solution (the stop solution is, for example, a new complete culture medium) for standby use.

[0100] S7. The control system is used to analyze the cell rounding rate in the real-time cell image data at equal intervals or in real time, and display and store the cell rounding rate that changes with time on a display screen connected to the control system. When the cell rounding rate reaches the designed value, the return mechanism 8 is started to return the culture dish from the integrated cell counting and cell morphology observation device 7 to the input and output mechanism 2.

[0101] S8. The cover opening mechanism 3 opens the returned culture dish, and the pipetting mechanism 1 adds the prepared stop solution into the culture dish.

[0102] In step S8, after the stop solution is added, the tilting mechanism 4 is controlled to tilt the culture dish again, and then the pipetting mechanism 1 is controlled to move, so that the pipette gun 125 sucks the stop solution on the culture dish and squeezes the stop solution onto the bottom plate of the tilted culture dish. The operation is repeated many times to simulate manual operation of blowing the cells on the bottom of the culture dish, so as to facilitate the rounded cells to fall off from the adherent state.

[0103] S9, the tube clamp 52 and tube cap opener 53 of the material preparation mechanism 5 cooperate to open a new test tube 54, and the new test tube 54 is clamped and supported by the tube clamp 52. Then the pipetting mechanism 1 is controlled to transfer all the liquid in the culture dish in S8 to the new test tube 54, and then the tube cap opener 53 is controlled to cover the tube cap of the new test tube 54 (after the tube cap is covered, the tube clamp 52 releases the clamp on the test tube 54), and the tube cap opener 53 is used to clamp the new test tube 54 and place the new test tube 54 on the centrifuge mechanism 6 for centrifugation.

[0104] In this step, the empty culture dish after the liquid is transferred is transported in the reverse direction by the input and output mechanism 2 , and the empty culture dish is sent back to the loading and unloading station, and then taken away by the loading and unloading manipulator 21 of the input and output mechanism 2 .

[0105] S10. The centrifuged test tube 54 is placed back on the spare rack 51, and the tube clamp 52 and the tube cap opener 53 are used to open the centrifuged test tube 54. The pipetting mechanism 1 sucks away the supernatant of the centrifuged test tube 54. Then, the pipetting mechanism 1 uses a new gun tip to inject new complete culture medium into the centrifuged test tube 54 to resuspend the centrifuged cells.

[0106] S11, the loading and unloading robot 21 takes the cell counting plate and places it on the loading and unloading turntable 22, and then the loading and unloading turntable 22 transfers the cell counting plate to the transfer station, and the conveyor belt 23 sends the cell counting plate to the bottom of the pipetting mechanism 1. The pipetting mechanism 1 takes the resuspended cell solution and injects the cell solution into the cell counting plate.

[0107] S12, with the cooperation of the receiving conveyor belt 23 and the pusher 24, the cell counting plate is sent to the integrated cell counting and cell morphology observation device 7 for cell counting. After the cell counting is completed, the control system actively records and stores the counting results, and controls the return mechanism 8 to send the cell counting plate back to the input and output mechanism 2. The input and output mechanism 2 sends the cell counting plate back to the loading and unloading station, and the loading and unloading robot 21 takes the cell counting plate away.

[0108] S13. After the cell counting is completed, the loading and unloading robot 21 sends the culture dishes to be used for subculture to the loading and unloading turntable 22 one by one, so that the new culture dishes to be used for subculture are sent to the receiving conveyor belt 23 in sequence.

[0109] S14. Inject the cell suspension and new complete culture medium into the new culture dish: the lid opening mechanism 3 opens the lid of the new culture dish, and the pipetting mechanism 1 adds the resuspended cell liquid into the new culture dish according to the subculture requirements. The control system controls the amount of cell suspension and new complete culture medium added to each new culture dish according to the cell counting results and subculture requirements.

[0110] S15, the cover opening mechanism 3 covers the lid of the new culture dish and uses the cover opening mechanism 3 to drive the new culture dish to move in a cross shape on the receiving conveyor belt 23, so that the cell fluid is evenly spread in the culture dish.

[0111] S16, the cover opening mechanism 3 moves away from the new culture dish, controls the input and output mechanism 2 to start in reverse, and sends the new culture dish back to the loading and unloading station in turn, and the loading and unloading robot 21 transfers the new culture dish sent back to the loading and unloading mechanism in turn to the designated position to complete the cell passage.

[0112] In this embodiment, the entire cell passaging process is fully automated through the provision of a cell passaging device. During the entire process, the operator only needs to prepare all the raw materials required for passaging in advance, place all the raw materials in the set positions, and mark them. In addition, the automated control system on the cell passaging device can automatically complete the fully automated operation, greatly reducing the difficulty of cell passaging and freeing up the hands of scientific researchers.

[0113] In addition, in this embodiment, the setting of the pipetting mechanism 1 can ensure that the pipetting process does not cause cross contamination between the pipetted liquids, and the setting of multiple pipette guns 125 can make the application range of cell passaging wider, and the entire passaging process can completely simulate manual operation.

[0114] In addition, this embodiment provides the input and output mechanism 2, the integrated cell counting and cell morphology observation device 7, and the return mechanism 8, so that both cell morphology observation and cell counting can be automated, and centrifugation can also be automatically performed during cell passage.

[0115] The above is only an embodiment of the present invention, and the common knowledge such as the specific technical solutions and / or characteristics in the solution are not described in detail here. It should be pointed out that for those skilled in the art, without departing from the technical solution of the present invention, several variations and improvements can be made, which should also be regarded as the scope of protection of the present invention, and these will not affect the effect of the implementation of the present invention and the practicality of the patent. The scope of protection required by this application shall be based on the content of its claims, and the specific implementation methods and other records in the description can be used to interpret the content of the claims.

Claims

1. A cell passage device comprising a pipetting mechanism, a material standby mechanism, and a lid opening mechanism. The pipetting mechanism comprises a spatial mover and a pipetting assembly mounted at an output end of the spatial mover, the spatial mover being used to drive the pipetting assembly to move in space; the material standby mechanism being used to place test tubes, open test tubes containing liquid raw materials, and open spare test tubes; and the lid opening mechanism being used to open and close the lid of a culture dish. The device is characterized by: The pipette assembly includes a presser, a rotating seat and multiple pipettes. The rotating seat and the presser are both installed at the output end of the space mover. The multiple pipettes are distributed circumferentially on the rotating seat. The rotating seat is used to rotate the pipette to the working position. The presser is used to press the pressing head of the pipette located in the working position to absorb or discharge liquid. The presser is used to press the pushing head rod after aligning the pushing head rod of the pipette to replace the gun head; the pipette mechanism also includes a waste container located below the pipette and a gun head group with multiple gun heads of different specifications. The pipette can be inserted into the gun head of the gun head group to assemble a new gun head. The gun head or waste liquid pushed out of the pipette can be received by the waste container.

2. A cell passaging device according to claim 1, characterized in that: The presser includes a reciprocating rod, a rotator and a pressing block. The reciprocating rod is used to drive the pressing block away from or close to the pressing head of the pipette gun. The pressing block and the pressing head can fit in a concave-convex manner. The rotator is used to drive the pressing block to rotate.

3. A cell passage device according to claim 2, characterized in that: The rotating seat is provided with a plurality of suction volume data acquisition modules, and each suction volume data acquisition module is used to acquire the measurement range of a corresponding pipette.

4. A cell passage device according to claim 3, characterized in that: A hollow cavity is provided in the middle of the rotating seat, and the pipette gun is installed on the four sides of the rotating seat. The axial ends of the hollow cavity are provided with through holes. The pipetting assembly also includes a connecting rod fixedly installed on the output end of the space mover, and the free end of the connecting rod is fixed with an end plate, and a visual module is installed on the end plate. The visual module is used to collect image data below the pipetting assembly.

5. A cell passaging device according to any one of claims 1 to 4, characterized in that: The pipetting assembly also includes multiple pushers, each pipette is correspondingly installed on the output end of a pusher, and the pusher is installed on the rotating seat. The pusher is used to control the pipette to approach or move away from the object to be sucked when the rotating seat rotates the pipette to the working position.

6. A cell passaging device according to any one of claims 1 to 4, characterized in that: It also includes a centrifugal mechanism. The material standby mechanism includes a standby rack, a tube clamp and a tube cap opener. The standby rack can rotate and is provided with multiple test tube placement holes. Test tubes with tube caps can be placed on the placement holes of the standby rack and the test tubes are supported by the standby rack. The tube clamp is used to clamp the test tube body rotated to the cap opening station, and the tube cap opener is used to clamp and twist open the tube cap of the test tube at the cap opening station. The space mover is used to drive the pipette gun of the pipetting assembly to extend into the test tube at the cap opening station to aspirate or inject liquid; the tube cap opener is also used to transfer the test tube between the standby rack and the centrifugal mechanism.

7. A cell passaging device according to any one of claims 1 to 4, characterized in that: The cover opening mechanism includes a multi-axis motion actuator and a suction cup installed at the output end of the multi-axis motion actuator. The multi-axis motion actuator is used to drive the suction cup to move in space, and the spatial movement includes movement of the X-axis, Y-axis, and Z-axis.

8. A cell passaging device according to any one of claims 1 to 5, characterized in that: It also includes an input and output mechanism, which is used to send culture dishes / cell counting plates into or out of the transfer operation area; the input and output mechanism includes an in-and-out material robot, an in-and-out material turntable and a receiving conveyor belt, the in-and-out material turntable includes a fixed tray and a rotatable turntable, the in-and-out material robot is used to take and place culture dishes / cell counting plates on the tray, the part of the tray facing the receiving conveyor belt is provided with a through hole, the turntable is used to switch the culture dishes / cell counting plates on the tray and the receiving conveyor belt in a rotating manner, and the receiving conveyor belt is used to transport the culture dishes to the bottom of the pipetting mechanism.

9. The cell passaging device according to claim 8, characterized in that: It also includes a return mechanism and an integrated cell counting and cell morphology observation device. The integrated cell counting and morphology observation device can not only count cells, but also observe and record the cell morphology in the culture dish. The integrated cell counting and morphology observation device is provided with a cell image data acquisition module, which is used to collect cell image data displayed on the integrated cell counting and morphology observation device; the input and output mechanism is provided with a pusher, which is used to send the culture dish / cell counting plate on the conveyor belt to the loading platform of the integrated cell counting and morphology observation device; the return mechanism is used to return the culture dish / cell counting plate on the loading platform to the input and output mechanism.

10. The cell passaging device according to claim 8, characterized in that: The receiving conveyor belt includes synchronous, parallel, and equal-height conveyor belts. There is a distance between the two conveyor belts. A tilting mechanism is provided between the two conveyor belts. The tilting mechanism includes a pusher and a push rod and a baffle fixed at the output end of the pusher. The push rod is used to push the culture dish. The distance between the push rod and the baffle is greater than the radius of the culture dish and smaller than the diameter of the culture dish. The baffle is used to allow the culture dish to tilt against the baffle when the push rod eccentrically pushes the culture dish.

Images