Cell counting and cell morphology observation integrated equipment

Abstract

The invention relates to the technical field of biological cell counting, and particularly discloses cell counting and cell morphology observation integrated equipment, which comprises a cell counting instrument capable of performing cell counting and cell morphology observation, and further comprises an image data acquisition module and a control module, the image data acquisition module is used for acquiring cell image data obtained after focusing of the cell counter in real time, the image data acquisition module is used for transmitting the cell image data to the control module, and the control module is used for analyzing the cell roundness rate in the cell image data and displaying and recording the cell roundness rate changing along with time. The scheme is used for solving the problem of poor cell passage consistency caused by inconsistent cell morphology judgment due to the fact that existing cell morphology observation depends on artificial experience judgment.

Description

Technical Field

[0001] The present invention relates to the technical field of biological cell counting, and in particular to an integrated device for cell counting and cell morphology observation. Background Art

[0002] In existing cell processing, microscopic equipment is often required to observe cell morphology and count cells.

[0003] Cell morphology observation, such as when passaging cells, requires examining the initial cell morphology under a microscope before passaging. The old complete medium in the culture dish is then discarded, and dead cell debris and residual complete medium are washed away with PBS. Digestive enzymes are then added, and the culture dish is placed under a microscope to observe morphology. Once the cells have grown to the desired state, fresh complete medium is added to the dish to terminate digestion. This process requires covering the culture dish twice with the lid and placing it on the stage under the microscope for observation of cell morphology.

[0004] Cell counting is mostly done during cell passaging and cell packaging, where the number of cells in the cell fluid needs to be calculated based on the passaging concentration and the packaging concentration. When counting, a certain amount of cell fluid is usually injected into a cell counting plate, and then the cell counting plate is placed on the stage of a microscope. Simple microscopes require manual counting, but many devices now have automatic counting functions. When the cell counting plate is placed on the microscope, the device will automatically focus and count.

[0005] Because both observing morphology and counting cells with a microscope utilizes the image acquisition function of the microscope, the prior art has proposed an integrated device that can both observe cell morphology and count cells. For example, patent publication number CN112577883A describes a small automated cell counter that can automatically count cells and observe cell morphology on a single device.

[0006] However, when observing the morphology of cells with this integrated cell counter, manual inspection is still required to determine the cell morphology. However, when manually inspecting the cell growth morphology, there are differences in the judgment of the cell growth status, resulting in large differences in the final results of the same cell culture when different people perform the same culture. Summary of the Invention

[0007] The present invention aims to provide an integrated device for cell counting and cell morphology observation to solve the problem of poor consistency in cell passage caused by inconsistent cell morphology judgments in existing cell morphology observations that rely on manual experience.

[0008] In order to achieve the above object, the present invention adopts the following technical solutions: A device integrating cell counting and cell morphology observation includes a cell counter capable of performing cell counting and cell morphology observation, an image data acquisition module, and a control module. The image data acquisition module is used to acquire cell image data obtained after focusing of the cell counter in real time, and the image data acquisition module is used to transmit the cell image data to the control module. The control module is used to analyze the cell rounding rate in the cell image data and display and record the cell rounding rate changing over time.

[0009] The principle and advantage of this solution are as follows: when using this solution, because the control module can analyze the real-time received cell image data to clearly display the cell rounding rate, the operator only needs to add the stop solution (such as complete culture medium) at the set rounding rate, which can greatly improve the consistency of the addition of the stop solution by different operators, thereby improving the consistency of cell passaging.

[0010] In addition, the use of a cell counter with an image data acquisition module and a control module can record the cell morphology changes in real time, which helps to record the research on different cells in scientific research.

[0011] Preferably, as an improvement, the control module is provided with a marking submodule, which is used to mark the cells confirmed to be rounded when calculating the cell rounding rate, so that the calculation of the cell rounding rate can be clearly recorded.

[0012] Preferably, as an improvement, the control module also includes a manual revision submodule, which is used to correct the rounded cells marked by the marking submodule, and the correction includes increasing or decreasing the marked rounded cells. The control module is used to recalculate the cell rounding rate after the manual revision submodule completes the correction. This solution can improve the control module's judgment accuracy on the cell rounding rate and improve flexibility through human-computer interaction.

[0013] Preferably, as an improvement, it also includes a pipetting device, which 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 pipetting assembly includes a presser, a rotating seat and multiple pipette guns, the rotating seat and the presser are both installed at the output end of the space mover, the multiple pipette guns are distributed circumferentially on the rotating seat, the rotating seat is used to rotate the pipette gun to the working position, the presser is used to press the pipette gun at the working position to absorb liquid or discharge liquid, and after the cell rounding rate obtained by analysis reaches the set value, the control module controls the pipette gun of the pipetting device to absorb the stop liquid and add it to the cell sample whose cell rounding rate reaches the set value.

[0014] When adopting this scheme, by setting up the pipetting device, when the cell growth needs to be stopped, the control module can automatically control the pipetting device to add the stop solution to the cell sample (such as the culture dish) within the set time, further ensuring the consistency of the passage.

[0015] In addition, the multiple pipettes provided with the pipette assembly can be easily installed with a variety of pipettes of different ranges to meet different usage requirements.

[0016] In addition, because the pipette has a push rod for replacing the pipette tip in addition to the pressing head for aspirating liquid, every time the pipette tip needs to be replaced, it is only necessary to control the rotating seat to rotate so that the pressing head is aligned with the push rod, and the used pipette tip can be automatically pushed out, thereby improving the degree of automation of the pipette device.

[0017] 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.

[0018] 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 for suction control of the pipette, the pressing block and the pressing head of the pipette 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 suction amount of the pipette is adjusted by screwing the pressing head with the pressing block.

[0019] Preferably, as an improvement, the pipetting device is further provided with a suction volume data acquisition module for capturing images of the suction volume of the pipette, and the control module is used to control the rotator to stop rotating the pressing block after the suction volume data of the corresponding pipette reaches a set value, so as to realize automatic adjustment and control of the suction volume of the pipette and improve the degree of automation.

[0020] Preferably, as an improvement, it further includes a pushing mechanism, a return mechanism and a cover opening mechanism. The pushing mechanism is located below the pipetting device. The pushing mechanism is used to push the cell sample onto the stage of the cell counter. The return mechanism includes a return device and a push-back plate. The return device is used to drive the push-back plate away from or close to the pushing mechanism. The return mechanism is used to push the cell sample on the stage back to the pushing mechanism; the cover opening mechanism is used to open or cover the cover of the cell sample.

[0021] When adopting this scheme, if the stop solution needs to be added to the culture dish serving as the cell sample, the return mechanism is first used to push the culture dish on the stage to the pushing mechanism. Driven by the pushing mechanism, the culture dish is transported to an area convenient for the opening of the cover by the opening mechanism and the transfer of liquid by the pipetting device. After the culture dish is opened, the pipette gun of the pipetting device adds the required liquid to the culture dish. The entire scheme realizes a fully automated process of automatic entry into the cell counter, automatic delivery out of the cell counter, automatic filling of liquid, and automatic opening of the cover, which facilitates the automated operation of cell morphology observation and the automated operation of cell counting.

[0022] Preferably, as an improvement, the cover opening mechanism and the pipetting device are located in the clean bench, the cell counter is located in the auxiliary detection room, the auxiliary detection room is connected to the clean bench, the clean bench is provided with a fan system, the fan system includes a filter, the fan system is used to provide air of set cleanliness to the clean bench, the pressure in the clean bench is greater than the pressure in the auxiliary detection room, the auxiliary detection room is provided with a pressure relief hole to ensure that the cleanliness in the clean bench is not affected by the auxiliary detection room, and at the same time, because the pressure in the clean bench is greater than the pressure in the auxiliary detection room, it is convenient to continuously send the clean air of the clean bench to the auxiliary detection room.

[0023] Preferably, as an improvement, the pushing mechanism includes a conveying part and a pushing part, the conveying part includes two synchronous, equal-height and parallel first conveyor belts, the pushing part is used to push the cell samples on the first conveyor belts from the clean bench to the loading platform; a tilting mechanism is also provided between the two first 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, so as to facilitate the pipette to absorb the liquid in the culture dish. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a flow chart of embodiment 1 of the present invention.

[0025] Figure 2 This is a schematic diagram of the three-dimensional structure of the second embodiment of the present invention.

[0026] Figure 3 for Figure 2 The top of the clean bench and auxiliary inspection room were removed to facilitate viewing of the three-dimensional schematic diagram of the internal structure.

[0027] Figure 4 for Figure 3 Schematic diagram of the three-dimensional structure after rotation angle.

[0028] Figure 5 For the general Figure 4Schematic diagram of the three-dimensional structure after the surrounding areas of the auxiliary inspection room and the transparent plate in front of the clean bench are removed.

[0029] Figure 6 for Figure 5 main view.

[0030] Figure 7 for Figure 5 Schematic diagram of the three-dimensional structure of the clean bench after the front, left side and back are removed (to show the positional relationship between the pipetting device, pushing mechanism, return mechanism and cell counter).

[0031] Figure 8 for Figure 7 Schematic diagram of the pipetting device in .

[0032] Figure 9 for Figure 8 Schematic diagram of the three-dimensional structure of the pipetting assembly when the reciprocating pressure rod moves downward to the pressing block and the pressing head of the pipette gun is concave and convex.

[0033] Figure 10 for Figure 9 main view.

[0034] Figure 11 for Figure 9 The three-dimensional schematic diagram after the mounting base is removed (a schematic diagram showing that the reciprocating pressure rod of the rack structure is driven by a motor to rotate the gear, and then the gear drives the reciprocating movement of the gear).

[0035] Figure 12 Figure 11 Bottom view of .

[0036] Figure 13 for Figure 7 Schematic diagram of the local three-dimensional structure after the pipetting device is not shown.

[0037] Figure 14 for Figure 13 main view.

[0038] Figure 15 for Figure 13 A top view of the first conveyor belt and the tilting mechanism is shown in FIG.

[0039] Figure 16 Schematic diagram of the three-dimensional structure of the tilting mechanism in the second embodiment of the present invention.

[0040] Figure 17 This is a front view of the process in which the tilting mechanism in Example 2 of the present invention pushes the culture dish to an inclined position (in order to conveniently illustrate the action process of the tilting mechanism, only the first conveyor belt is shown which does not block the front view of the tilting mechanism). DETAILED DESCRIPTION

[0041] The following is further described in detail through specific implementation methods: The figure marks in the drawings of the specification include: cell counter 10, stage 101, clean bench 20, auxiliary detection chamber 30, pipetting device 1, space mover 11, pipetting assembly 12, mounting seat 121, presser 122, pressing drive source 1221, reciprocating pressure rod 1222, rotator 1223, pressing block 1224, rotating seat 123, pusher 124, pipette gun 125, pressing head 1251, pushing head rod 1252, suction volume data acquisition module 126, pushing mechanism 2, first conveyor belt 21, second conveyor belt 22, pushing plate 221, transition support platform 23, cover opening mechanism 3, multi-axis motion actuator 31, suction cup 32, tilting mechanism 4, pusher 41, push rod 42, baffle 43, return mechanism 5, returner 51, push back plate 52, culture dish 100.

[0042] Example 1 Combine Figure 1 A device integrating cell counting and cell morphology observation includes a cell counter 10. The cell counter 10 adopts an existing cell counter 10 with cell counting and cell morphology observation functions. The cell counter 10 has an automatic focusing function and can automatically adjust the focus according to the cell morphology to be observed or the cell count to be performed (the cell morphology to be observed or the cell count to be performed is collectively referred to as a cell sample).

[0043] It also includes an image data acquisition module and a control module. The image data acquisition module is used to collect clear cell image data obtained after the cell counter 10 is focused in real time. The image data acquisition module is used to transmit the collected real-time cell picture data to the control module. The control module includes analysis mode selection, and the analysis mode selection includes morphological observation and cell counting. The control module is used to count the total number of cells in the cell counting plate after the analysis mode is selected as cell counting. The control module is used to calculate the cell rounding rate of the cells in the culture dish after the analysis mode is selected as morphological observation.

[0044] The control module 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 module (display / store a curve of the cell rounding rate over time). The image data acquisition module can use a CCD camera.

[0045] The control module includes a marking submodule, which marks cells that have been confirmed to be rounded when calculating the cell rounding rate. This ensures that the calculated cell rounding rate can be clearly stored and recorded. The operator can access and review the cell rounding rate at a specific time on the display, along with the cell marking status of the marking submodule at that time.

[0046] The control module also includes a manual revision submodule, which is used to correct the rounded cells marked by the marking submodule. The correction includes increasing or decreasing the marked rounded cells (adding marks: for example, allowing users to select unidentified rounded cells; deleting marks: clearing misjudged cell marks, such as bubbles and cell debris).

[0047] The control module is used to recalculate the cell rounding rate after the manual revision submodule completes the correction. The control module performs machine learning on the data revised by the manual revision submodule. For example, the control module extracts the morphological features of the corrected rounded cells (morphological features include area, circularity, and edge integrity). Based on the extracted morphological features of the rounded cells, the control module updates the machine learning model used to determine cell roundness (e.g., using a random forest / SVM algorithm). This allows the control module to continuously improve the accuracy of cell rounding rate judgment through human-computer interaction, increasing flexibility.

[0048] The cell rounding rate = the number of cells judged as "round" / the total number of valid cells.

[0049] When the circularity, area, and edge integrity of a cell reach the set values, the cell is judged as a "round" cell. The circularity reaches the set value because the vast majority of cells are adherent cells. During the growth process of adherent cells, the shape gradually changes from spindle to round and gradually detaches from the adherent state. The area limitation is to avoid incorporating bubbles or cell fragments into round cells, and the edge integrity is to eliminate cells that are round but damaged.

[0050] When using this embodiment, since the control module can analyze the cell image data received in real time to clearly display the cell rounding rate, the operator only needs to add the stop solution (such as complete culture medium) at the set rounding rate to greatly improve the consistency of the addition of the stop solution by different operators, thereby improving the consistency of cell passaging.

[0051] In addition, the use of the cell counter 10 in conjunction with the image data acquisition module and the control module can record the cell morphology change process in real time, which is helpful for recording the research of different cells in scientific research.

[0052] In addition, the control module can also calculate the cell rounding rate in real time. Researchers can explore the effects of adding stop solution at different intervals on cells, making scientific research records simpler and more convenient, and facilitating more comparative experiments.

[0053] Example 2 The second embodiment is improved on the basis of the first embodiment as follows: Combine Figures 2 to 17 A cell counting and cell morphology observation integrated device also includes a clean bench 20 and an auxiliary detection room 30. The clean bench 20 is connected to the auxiliary detection room 30. The cell counter 10 is placed in the auxiliary detection room 30. The clean bench 20 is provided with a fan system. The fan system includes a filter. The fan system is used to provide air with a set cleanliness level to the clean bench 20. The pressure in the clean bench 20 is greater than the pressure in the auxiliary detection room 30. The auxiliary detection room 30 is provided with a pressure relief hole to ensure that the cleanliness level in the clean bench 20 is not affected by the auxiliary detection room 30. At the same time, because the pressure in the clean bench 20 is greater than the pressure in the auxiliary detection room 30, it is convenient to continuously send the clean air in the clean bench 20 to the auxiliary detection room 30.

[0054] The sides and top of the auxiliary detection chamber 30 are transparent, so that the staff can directly see the detection status of the cell counter 10 through the auxiliary detection chamber 30.

[0055] The clean bench 20 is provided with a pipetting device 1, a pushing mechanism 2, a lid opening mechanism 3, and a tilting mechanism 4, and the auxiliary detection chamber 30 is provided with a return mechanism 5. The pipetting device 1 is used to add or extract liquid from a culture dish placed on the pushing mechanism 2, and the pipetting device 1 is also used to inject the cell fluid to be tested into a cell counting plate placed on the pushing mechanism 2; the pushing mechanism 2 is used to support the culture dish and / or cell counting plate, and is used to push the culture dish / cell counting plate from the clean bench 20 through the connecting channel between the clean bench 20 and the auxiliary detection chamber 30 to the stage 101 on the cell counter 10. After the culture dish or cell counting plate is pushed onto the stage 101, the cell counter 10 can observe the cell morphology or count the cells on the culture dish or cell counting plate.

[0056] The cover opening mechanism 3 is used to open the cover of the culture dish placed on the pushing mechanism 2 to facilitate the pipetting device 1 to add or extract liquid into or out of the culture dish.

[0057] The tilting mechanism 4 is used to tilt the circular culture dish to a tilted state, so that the pipetting device 1 can more easily suck the liquid in the culture dish.

[0058] The return mechanism 5 is used to push the culture dish / cell counting plate on the stage 101 after completing cell counting or morphological observation back to the pushing mechanism 2, so as to facilitate the culture dish / cell counting plate to be returned to the clean bench 20 after completing morphological observation or cell counting.

[0059] The specific structure is as follows: 1. Liquid handling device 1 Combine Figures 8 to 12 The pipetting device 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, 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 device 1.

[0060] The pipetting assembly 12 includes a mounting base 121, a presser 122, a rotating base 123, a plurality of pushers 124 and a plurality of pipette guns 125. The mounting base 121 is fixed to the output end of the space mover 11. The rotating base 123 and the presser 122 are both mounted on the mounting base 121. The plurality of pushers 124 are evenly distributed circumferentially around the rotating structure of the rotating base 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 mounted on the output end of each pusher 124. The rotating seat 123 is used to rotate the pipette 125 to the working position, and the pusher 124 is used to control the pipette 125 to move closer to or away from the object to be sucked when the rotating seat 123 rotates the pipette 125 to the working position. The presser 122 is used to press the pipette 125 located in the working position to absorb liquid or discharge liquid. After the cell rounding rate obtained by analysis reaches the set value, the control module controls the pipette 125 of the pipetting device 1 to absorb the stop liquid and add it to the cell sample whose cell rounding rate reaches the set value.

[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. The output slider is detachably connected to a pipette 125. The pipette 125 can be a pipette 125 disclosed in patent publication number CN213193738U.

[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 push rod 1252 for replacing the pipette tip in addition to the pressing head 1251 for aspirating liquid, each time the pipette tip 125 needs to be replaced, it is only necessary to control the rotating seat 123 to rotate so that the pressing head 1251 is aligned with the push rod 1252, and the used pipette tip 125 can be automatically pushed out, thereby improving the automation level of the pipetting device 1. The pipette 125 of this embodiment can adopt the pipette 125 disclosed in patent publication number CN213193738U.

[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.

[0066] The control module is used to control the rotator 1223 to stop rotating the pressing block 1224 after the suction volume data of the corresponding pipette 125 reaches a set value, so as to realize automatic adjustment and control of the suction volume of the pipette 125 and improve the degree of automation.

[0067] 2. Opening mechanism 3 Combine Figures 13 and 14 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 used to drive the suction cup 32 to move in 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 device 1 to add or absorb liquid to the culture dish.

[0068] 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.

[0069] 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.

[0070] In addition, the suction cup 32 of this embodiment is an elastic suction cup 32, so that when the multi-axis motion actuator 31 drives the suction cup 32 to press down on the lid of the culture dish, there is elastic buffering, avoiding the problem that the multi-axis motion actuator 31 presses down a slightly larger distance and crushes or damages the culture dish.

[0071] 3. Pushing mechanism 2, return mechanism 5 and tilting mechanism 4 Combine Figures 13 to 17 The pushing mechanism 2 is located below the pipetting device 1 , and is used to push the cell sample onto the stage 101 of the cell counter 10 .

[0072] The pushing mechanism 2 includes a conveying part and a pushing part. The conveying part is installed in the ultra-quiet table. The conveying part includes two synchronous and parallel first conveyor belts 21. The two first conveyor belts 21 have the same height and the same conveying direction. The pushing part includes a second conveyor belt 22. The second conveyor belt 22 spans the ultra-clean table 20 and the auxiliary detection room 30. The second conveyor belt 22 is located between the two first conveyor belts 21 and above the end of the first conveyor belt 21 near the cell counter 10. The height of the first conveyor belt 21 is the same as the height of the stage 101 of the cell counter 10. The ultra-clean table 20 and the auxiliary detection room The second conveyor belt 22 is provided with a transition support platform 23. The transition support platform 23 is at the same height as the first conveyor belt 21 and is located between the loading platform 101 and the first conveyor belt 21. A push plate 221 is fixed on the second conveyor belt 22. After the second conveyor belt 22 is started, the push plate 221 is used to push the cell sample (cell sample, i.e., culture dish / cell counting plate) on the first conveyor belt 21 through the transition support platform 23 and onto the loading platform 101, making it convenient to send the cell sample in the clean bench 20 to the cell counter 10 in the auxiliary detection room 30 for cell morphology observation or cell counting. The clean bench 20 and the auxiliary detection room 30 are both provided with windows for convenient placement of culture dishes and cell counting plates.

[0073] The return mechanism 5 includes a return device 51 and a return push plate 52. The return device 51 is used to drive the return push plate 52 away from or close to the push mechanism 2. In this embodiment, the return device 51 adopts a linear motor or a cylinder. The return mechanism 5 is used to push the cell sample on the stage 101 back to the push mechanism 2. In this embodiment, after the cell sample morphology observation or cell counting is completed, the return mechanism 5 installed in the auxiliary detection chamber 30 pushes the cell sample back to the first conveyor belt 21 or the transition support table 23. If it is pushed back to the transition support table 23, the reverse conveyance of the second conveyor belt 22 is utilized to push the cell sample to the first conveyor belt 21 under the push of the push plate 221. If the return mechanism 5 has already pushed the cell sample to the first conveyor belt 21, the reverse conveyance of the first conveyor belt 21 can be directly controlled to allow the cell sample to be returned to the clean bench 20 again.

[0074] A tilting mechanism 4 is also provided between the two first conveyor belts 21. The 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 41 drives the push rod 42 and the baffle 43 to rise and fall synchronously along the Z axis. The pusher 42 is an elastic push rod 42. The push rod 42 is used to push the culture dish. When pushing the culture dish, the elastic push rod 42 uses its elasticity to slowly push the culture dish to avoid overturning the culture dish by pushing too quickly; the distance between the push rod 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 push rod 42 eccentrically pushes the culture dish, so as to facilitate the pipette gun 125 to absorb the liquid in the culture dish. In this embodiment, the baffle 43 is an arc-shaped plate, which cooperates with the arc structure of the culture dish to further ensure that after the push rod 42 pushes the culture dish to tilt, the culture dish will not tip over in other directions.

[0075] The tilting mechanism 4 is designed because during cell passaging, a series of operations need to be performed on the cells in the culture dish before morphological observation. For example, the old culture medium in the culture dish to be passaged must first be completely removed. If the culture dish is placed flat on the first conveyor belt 21, it is difficult to ensure that the old culture medium is fully extracted. The upward push of the tilting mechanism 4 simulates the manual tilting of a conventional circular culture dish. Similarly, after the cell morphological observation is completed, when the cell fluid in the culture dish needs to be completely transferred to a centrifuge tube, the culture dish also needs to be tilted to ensure that the cell fluid can be basically completely extracted.

[0076] Therefore, this embodiment, through the arrangement of the pipetting device 1, the cover opening mechanism 3, the pushing mechanism 2, the return mechanism 5 and the tilting mechanism 4, can realize the automation of liquid filling and removal, the automation of culture dish opening, and the automation of cell sample inspection during cell passaging, greatly improving the automation level of cell processing, helping to reduce the tedious manual operations of scientific researchers, ensuring the consistency of cell processing, and saving more time for scientific researchers.

[0077] 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. An integrated device for cell counting and cell morphology observation, comprising a cell counter capable of performing cell counting and cell morphology observation, characterized in that: It also includes an image data acquisition module and a control module. The image data acquisition module is used to collect cell image data obtained after the cell counter is focused in real time. The image data acquisition module is used to transmit the cell image data to the control module. The control module is used to analyze the cell rounding rate in the cell image data and display and record the cell rounding rate that changes with time.

2. The integrated device for cell counting and cell morphology observation according to claim 1, characterized in that: The control module is provided with a marking submodule, and the marking submodule is used to mark the cells confirmed to be rounded when the cell rounding rate is calculated.

3. The integrated device for cell counting and cell morphology observation according to claim 2, characterized in that: The control module also includes a manual revision submodule, which is used to correct the rounded cells marked by the marking submodule, and the correction includes increasing or decreasing the marked rounded cells. The control module is used to recalculate the cell rounding rate after the manual revision submodule completes the correction.

4. The integrated cell counting and cell morphology observation device according to any one of claims 1 to 3, characterized in that: It also includes a pipetting device, which 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 pipetting 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 around the rotating seat. The rotating seat is used to rotate the pipette to the working position. The presser is used to press the pipette located at the working position to absorb or discharge liquid. After the cell rounding rate obtained by analysis reaches the set value, the control module controls the pipette of the pipetting device to absorb the stop liquid and add it to the cell sample whose cell rounding rate reaches the set value.

5. The integrated device for cell counting and cell morphology observation according to claim 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. The integrated device for cell counting and cell morphology observation according to claim 5, characterized in that: 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 suction control pressing head of the pipette. The pressing block and the pressing head of the pipette can fit together in a concave and convex manner. The rotator is used to drive the pressing block to rotate.

7. The integrated device for cell counting and cell morphology observation according to claim 6, characterized in that: The pipetting device is also provided with a suction volume data acquisition module for collecting images of the suction volume of the pipette, and the control module is used to control the rotator to stop rotating the pressing block after the suction volume data of the corresponding pipette reaches a set value.

8. The integrated device for cell counting and cell morphology observation according to claim 4, characterized in that: It also includes a pushing mechanism, a return mechanism and a cover opening mechanism. The pushing mechanism is located below the pipetting device. The pushing mechanism is used to push the cell sample onto the stage of the cell counter. The return mechanism includes a return device and a push-back plate. The return device is used to drive the push-back plate away from or close to the pushing mechanism. The return mechanism is used to push the cell sample on the stage back to the pushing mechanism; the cover opening mechanism is used to open or cover the cover of the cell sample.

9. The integrated device for cell counting and cell morphology observation according to claim 8, characterized in that: The opening mechanism and the pipetting device are located in the clean bench, the cell counter is located in the auxiliary detection room, the auxiliary detection room is connected to the clean bench, the clean bench is provided with a fan system, the fan system includes a filter, the fan system is used to provide air with a set cleanliness level into the clean bench, the pressure in the clean bench is greater than the pressure in the auxiliary detection room, and the auxiliary detection room is provided with a pressure relief hole.

10. The integrated cell counting and cell morphology observation device according to any one of claims 8 to 9, characterized in that: The pushing mechanism includes a conveying part and a pushing part. The conveying part includes two synchronous, equal-height and parallel first conveyor belts. The pushing part is used to push the cell samples on the first conveyor belts from the clean bench to the sample loading platform. A tilting mechanism is also provided between the two first 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.

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