Cell culture apparatus

Abstract

The invention relates to the technical field of life science instruments, and discloses cell culture equipment. A working area and a non-working area are arranged in an operation cabinet of the cell culture equipment, a liquid storage module is used for storing liquid at a preset storage temperature, and a liquid supply module is communicated with the liquid storage module and is used for heating a preset amount of liquid to a preset use temperature; the working area is further provided with a bearing seat, the pipetting module is provided with a pipetting head, the pipetting head is detachably arranged on the bearing seat, and the mechanical arm can selectively clamp and drive the pipetting head to carry out pipetting; the mechanical arm can move and operate between the pipetting module and the more than two operation modules; the waste area is selectively communicated with the working area; the culture cabinet and the consumable cabinet can be selectively communicated with the operation cabinet, the culture cabinet is used for cell culture, and the consumable cabinet is used for containing consumables. According to the invention, liquid required by cell culture can be stored and supplied according to different temperatures, meanwhile, the pipetting composition is more efficient and reliable, in addition, zoning is more reasonable, and the automation degree is higher.

Description

Technical Field

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

[0002] With the advancement of precision medicine, more and more research is conducted based on individual cells, such as antibody drug development, anti-cancer drug development, drug toxicological properties research, etc., so the demand for cell culture equipment has increased.

[0003] In the existing cell culture equipment, manual operation is the main method, which leads to the problems of poor consistency of cell culture between batches and inability to standardize the cell culture process. In addition, the storage and supply temperatures of various liquids required for cell culture are not suitable, which will cause damage to cell growth, and the pipetting process is prone to failure and low efficiency. In addition, the above problems will also lead to poor unattended operation capabilities of cell culture equipment.

[0004] Therefore, there is an urgent need for a cell culture device to solve the above problems. Summary of the invention

[0005] Based on the above, the purpose of the present invention is to provide a cell culture device that can store and supply liquids required for cell culture at different temperatures, while making the pipetting structure more efficient and reliable, and the partitioning more reasonable and the degree of automation higher.

[0006] To achieve the above object, the present invention adopts the following technical solutions:

[0007] Cell culture equipment, including:

[0008] An operating cabinet, wherein a working area and a non-working area are arranged in the operating cabinet, the working area is provided with a mechanical arm, a liquid supply module, a pipetting module and at least two operating modules, the non-working area is provided with a liquid storage module and a waste area, the liquid storage module is used to store liquid at a preset storage temperature, the liquid supply module is connected with the liquid storage module, and is used to heat a preset amount of liquid to a preset use temperature; the working area is also provided with a bearing seat, the pipetting module is provided with a pipetting head, the pipetting head is detachably arranged on the bearing seat, the mechanical arm can selectively clamp and drive the pipetting head to perform pipetting; the mechanical arm can move and operate between the pipetting module and more than two operating modules; the waste area is selectively connected with the working area;

[0009] The culture cabinet and the consumable cabinet are respectively arranged on both sides of the operation cabinet and can be selectively connected with the operation cabinet. The culture cabinet is used for cell culture, and the consumable cabinet is used for storing consumables.

[0010] As a preferred solution of the cell culture device, the pipetting module is further provided with a wire supply assembly, which includes a cable with adjustable length, one end of the cable is arranged on the supporting seat, and the free end is connected to the pipetting head.

[0011] As a preferred solution for the cell culture equipment, the liquid storage module includes a storage box, a plurality of liquid storage bottles and a detection unit. A refrigeration unit is arranged in the storage box, and the cold end of the refrigeration unit is connected to the inner cavity of the storage box so that the inner cavity has a preset storage temperature; a plurality of liquid storage bottles are arranged in the inner cavity, and the liquid storage bottles can be selectively connected to the main liquid pipe; a detection module is arranged in the inner cavity, and is arranged one-to-one with the liquid storage bottles to detect the remaining liquid in the liquid storage bottles.

[0012] As a preferred solution of the cell culture equipment, the liquid supply module includes a preheating unit, which includes a heating component and a consumable bag. The consumable bag includes a bag body, an outlet pipe and an inlet pipe. One end of the outlet pipe is connected to the bag body, and the other end is connected to the pipetting module. One end of the inlet pipe is connected to the bag body, and the other end is connected to the liquid storage module. The end of the inlet pipe away from the bag body is also connected to the air pipe, and the inlet pipe can selectively connect to the liquid storage module and the air pipe.

[0013] As a preferred solution of the cell culture equipment, an operating table is provided in the operating cabinet, and the operating table is used to separate the working area and the non-working area. A waste port and an isolation door are provided on the operating table, and the isolation door can selectively block the waste port to selectively connect the waste area and the working area.

[0014] As a preferred solution of the cell culture device, the waste area is also provided with a waste carrier, a waste switch door and a sliding assembly, the waste carrier is arranged on the sliding assembly, and the sliding assembly is used to move the waste carrier out of the waste switch door.

[0015] As a preferred solution of the cell culture device, the operation module includes a cover opening unit for opening the centrifuge tube and / or the culture bottle; and / or the operation module includes an incubation unit for providing a preset incubation temperature.

[0016] As a preferred solution of the cell culture equipment, the operating module also includes a filtering unit for filtering and circulating the air in the working area; and / or the operating module includes a scanning unit for scanning the characteristic code and / or characteristic chip of the culture bottle and / or centrifuge tube.

[0017] As a preferred solution of the cell culture equipment, a sterilization module is provided in the culture cabinet for providing hydrogen peroxide fumigation operation.

[0018] As a preferred solution of the cell culture device, the cell culture device further comprises a transport robot for transporting between the culture cabinet, the operation cabinet and the consumable cabinet.

[0019] The beneficial effects of the present invention are:

[0020] The present invention is provided with an operation cabinet, a culture cabinet and a consumable cabinet, which are respectively used for operation, culture and storage of consumables, and the culture cabinet and the consumable cabinet are respectively provided on both sides of the operation cabinet and can be selectively connected with the operation cabinet, so that the three cabinets can be connected or disconnected as needed, and are used to transfer materials when connected, and are used to maintain a relatively independent relationship when disconnected, so as to maintain a suitable environment for respective operations. A working area, a non-working area and a waste area are provided in the operation cabinet, and the zoning is more reasonable, reducing the influence between different modules in each area. The liquid storage module in the non-working area is used to store liquid at a preset storage temperature to ensure the activity and effectiveness of the liquid; the liquid supply module is used to heat a preset amount of liquid to a preset use temperature to ensure normal growth of cells; the liquid transfer head of the liquid transfer module is separated from the bearing seat under the drive of the mechanical arm to transfer liquid, and the liquid transfer process is more reliable and more efficient; at the same time, the mechanical arm can also move between multiple modules to perform corresponding operations, with good compatibility and high utilization rate. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments of the present invention. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the contents of the embodiments of the present invention and these drawings without paying any creative work.

[0022] Figure 1 is a schematic diagram of a cell culture device provided in a specific embodiment of the present invention;

[0023] Figure 2 It is a perspective schematic diagram of an operating cabinet of a cell culture device provided in a specific embodiment of the present invention;

[0024] Figure 3 is a schematic diagram of another perspective of the operating cabinet of the cell culture device provided by a specific embodiment of the present invention;

[0025] Figure 4 is a schematic diagram of the interior of an operating cabinet of a cell culture device provided in a specific embodiment of the present invention;

[0026] Figure 5 is a schematic diagram of another viewing angle of the operating cabinet of the cell culture device provided in a specific embodiment of the present invention;

[0027] Figure 6 is a schematic diagram of a pipetting module provided in a specific embodiment of the present invention;

[0028] Figure 7 It is a perspective schematic diagram of a pipetting component of a pipetting module provided in a specific embodiment of the present invention;

[0029] Figure 8 is a schematic diagram from another perspective of the pipetting component of the pipetting module provided in a specific embodiment of the present invention;

[0030] Fig. 9 It is a schematic diagram of the principle of a liquid storage module provided in a specific embodiment of the present invention;

[0031] Fig.10 is a schematic diagram of a liquid storage module provided in a specific embodiment of the present invention;

[0032] Fig.11 is a cross-sectional view of a liquid storage module provided in a specific embodiment of the present invention;

[0033] Fig.12 is a schematic diagram of a liquid supply module provided in a specific embodiment of the present invention;

[0034] Fig.13 is a cross-sectional view of a liquid supply module provided in a specific embodiment of the present invention;

[0035] Fig.14 It is a schematic diagram of a consumable bag of a liquid supply module provided in a specific embodiment of the present invention.

[0036] In the figure:

[0037] 1000, operation cabinet; 1001, work area; 1011, mechanical arm; 1016, pipetting head; 1017, filter unit; 1018, 96-channel pipetting device; 1019, information identification area; 1020, centrifuge tube consumables group; 1021, culture bottle consumables group; 1022, incubation unit; 1023, well plate nozzle box; 1024, pipetting nozzle box; 1025, cabinet consumables Transfer; 1026, disinfectant filling device; 1027, transfer during transfer; 1002, non-working area; 1100, rack; 1300, first window; 1400, second window; 1500, cover opening unit; 1510, electric suction cup cover opening machine; 1520, rotary cover opening machine; 1600, microscope; 1701, waste carrier; 1800, centrifuge; 1900, waste port;

[0038] 2000, culture cabinet;

[0039] 3000, consumables cabinet;

[0040] 4100, pipetting assembly; 4110, pipetting body; 4111, female snap-on connector; 4112, limit plate; 4114, male snap-on connector; 4300, clamping assembly; 4400, wire supply assembly; 4410, wire reel;

[0041] 5100, storage box; 5101, inner cavity; 5110, refrigeration unit; 5111, cold end; 5112, hot end; 5113, cooling fan; 5120, temperature averaging fan; 5131, inner layer; 5132, outer layer; 5133, insulation layer; 5200, liquid storage bottle; 5210, main liquid pipe; 5211, peristaltic pump; 5300, detection unit; 5310, weighing table; 5311, tray; 5312, weight sensor; 5400, preheating unit; 5500, control valve group; 5510, switch valve;

[0042] 6100, heating body; 6110, locking assembly; 6120, insulation layer; 6130, hinge; 6200, consumable bag; 6210, bag body; 6220, outlet pipe; 6230, inlet pipe; 6240, liquid pipe; 6250, air pipe; 6300, heating assembly; 6310, heating element; 6320, heat conductive block. DETAILED DESCRIPTION

[0043] Embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present invention, and should not be construed as limiting the present invention.

[0044] In the description of the present invention, it should be noted that the terms "center", "up", "down", "left", "right", "vertical", "horizontal", "inside", "outside" and the like indicate positions or positional relationships based on the positions or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, the terms "first" and "second" are used for descriptive purposes only and cannot be understood as indicating or implying relative importance. Among them, the terms "first position" and "second position" are two different positions.

[0045] Unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", and "fixed" should be understood in a broad sense, for example, it can be a fixed connection or a detachable connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0046] In the present invention, unless otherwise clearly specified and limited, a first feature being "above" or "below" a second feature may include that the first and second features are in direct contact, or may include that the first and second features are not in direct contact but are in contact through another feature between them. Moreover, a first feature being "above", "above" and "above" a second feature includes that the first feature is directly above and obliquely above the second feature, or simply indicates that the first feature is higher in level than the second feature. A first feature being "below", "below" and "below" a second feature includes that the first feature is directly below and obliquely below the second feature, or simply indicates that the first feature is lower in level than the second feature.

[0047] The technical solution of the present invention is further described below with reference to the accompanying drawings and through specific implementation methods.

[0048] like Figure 1-Figure 5 As shown, this embodiment provides a cell culture device, which includes an operation cabinet 1000, a culture cabinet 2000 and a consumables cabinet 3000. The operation cabinet 1000 is provided with a working area 1001 and a non-working area 1002. The working area 1001 is provided with a mechanical arm 1011, a liquid supply module, a pipetting module and at least two operation modules. The non-working area 1002 is provided with a liquid storage module and a waste area. The liquid storage module is used to store liquid at a preset storage temperature. The liquid supply module is connected to the liquid storage module and is used to heat a preset amount of liquid to a preset use temperature. The working area 1001 is provided with a mechanical arm 1011, a liquid supply module, a pipetting module and at least two operation modules. The non-working area 1002 is provided with a liquid storage module and a waste area. The liquid storage module is used to store liquid at a preset storage temperature. The liquid supply module is connected to the liquid storage module and is used to heat a preset amount of liquid to a preset use temperature. The 01 domain is also provided with a supporting seat, the pipetting module is provided with a pipetting head 1016, and the pipetting head 1016 is detachably arranged on the supporting seat, and the robotic arm 1011 can selectively clamp and drive the pipetting head 1016 for pipetting; the robotic arm 1011 can move and operate between the pipetting module and two or more operating modules; the waste area is selectively connected with the working area 1001; the culture cabinet 2000 and the consumable cabinet 3000 are respectively arranged on both sides of the operating cabinet 1000, and can be selectively connected with the operating cabinet 1000, the culture cabinet 2000 is used for cell culture, and the consumable cabinet 3000 is used for holding consumables.

[0049] By setting up an operation cabinet 1000, a culture cabinet 2000 and a consumable cabinet 3000, they are used for operation, culture and storage of consumables respectively, and the culture cabinet 2000 and the consumable cabinet 3000 are respectively set on both sides of the operation cabinet 1000, and can be selectively connected with the operation cabinet 1000, so that the three cabinets can be connected or disconnected as needed, and are used to transfer materials when connected, and are used to maintain a relatively independent relationship when disconnected, so as to maintain a suitable environment for separate operations. The operation cabinet 1000 is provided with a working area 1001, a non-working area 1002 and a waste area, and the zoning is more reasonable, reducing the impact between different modules in each area. The liquid storage module in the non-working area 1002 is used to store liquid at a preset storage temperature to ensure the activity and effectiveness of the liquid; the liquid supply module is used to heat a preset amount of liquid to a preset use temperature to ensure normal cell growth; the pipetting head 1016 of the pipetting module is separated from the supporting seat 1015 by the drive of the robotic arm 1011 to perform pipetting, and the pipetting process is more reliable and more efficient; at the same time, the robotic arm 1011 can also move between multiple modules to perform corresponding operations, with good compatibility and high utilization rate.

[0050] Specifically, the operation cabinet 1000, the culture cabinet 2000 and the consumable cabinet 3000 can be operated independently of each other, or can be dispatched and controlled through a bus. The three cabinets can be connected or independent through electrically controlled doors.

[0051] The culture cabinet 2000 can provide a suitable environment for the growth of cells, such as providing a suitable temperature field, a carbon dioxide field, etc. At the same time, a sterilization module is provided in the culture cabinet 2000 for providing a hydrogen peroxide fumigation operation. In addition, the culture cabinet 2000 can also be equipped with a set of motion mechanisms to transfer the cell culture containers inside the culture cabinet 2000, and can be transferred to a specific position. The culture cabinet 2000 is compatible with cell culture containers such as T75, well plates, and culture dishes. Optionally, the above-mentioned cell culture containers can be compatible with the motion mechanism through an intermediate conversion tray 5311, and the intermediate conversion tray 5311 can limit the freedom of the cell culture container in the plane direction. In particular, the culture cabinet 2000 can also be used for the culture of cells after filling with drugs or viruses.

[0052] The consumable cabinet 3000 can contain culture container consumables, orifice plate consumables, and disposable gun tips for pipetting. Optionally, the consumable cabinet 3000 is independently arranged outside the incubator; it can be contained in the operation cabinet 1000, or it can be independent of the operation cabinet 1000. Considering the working flux of the cell culture equipment, the consumable cabinet 3000 is preferably arranged outside the operation cabinet 1000. The consumable cabinet 3000 is also provided with a specific position to dock with the external automated feeding equipment. The specific position can be one or more. The specific position is set on the side of the consumable cabinet 3000, and the distance between the structural high point of the specific position and the ground is 800mm-1200mm. On the table of the work area 1001, there are arranged several work stations for placing consumables, and the wall of the work area 1001 is also hung with a station for placing consumables. For example, a centrifuge tube consumable group 1020, a culture bottle consumable group 1021, a orifice plate nozzle box 1023, and a pipetting nozzle box 1024.

[0053] Furthermore, the operation cabinet 1000 and the consumable cabinet 3000 are docked through an electric opening and closing door. The consumable cabinet 3000 is also provided with a device for transferring the feeding docking, and an automated electric control door, which is arranged on the side of the consumable cabinet 3000 close to the operation cabinet 1000, and is opened and closed in accordance with the needs of the device for transferring the feeding docking. The electric control door can be a swing-opening door or a flat-opening door. Preferably, a flat-opening door is adopted. The translational electric control door can be opened directly by a linear screw drive, or it can be opened and closed by a gear rack transmission.

[0054] The operation cabinet 1000 is provided with a rack 1100, a first window 1300 connected to the culture cabinet 2000, and a second window 1400 connected to the consumable cabinet 3000. The operation cabinet 1000 is divided into a working area 1001 and a non-working area 1002. The operation module also includes a filter unit 1017 for filtering and circulating the air in the working area 1001 to achieve circulating wind to ensure the cleanliness of the working area 1001. Optionally, a HEAP filter with high efficiency filtration is provided.

[0055] The operation module includes a cover opening unit 1500, which is used to open the centrifuge tube and / or culture bottle. One or more cover opening units 1500 are arranged in the specific working area 1001, some of the cover opening units 1500 are used for opening the centrifuge tube, and other parts of the cover opening units 1500 are used for opening the culture bottle. The number of the cover opening units 1500 for opening the centrifuge tube and the cover opening units 1500 for opening the culture bottle is 1:1, or 2:1. Specifically, the cover opening unit 1500 can realize the functions of Z-axis movement, local rotation and movement, so as to open the centrifuge tube and the culture bottle. The centrifuge tube and the culture bottle can be arranged in a well plate, and the well plate can be a 96-well plate or a 382-well plate, etc. Optionally, the cover opening unit 1500 includes an electric suction cup cover opening machine 1510 and a rotary cover opening machine 1520, which are used for different cover opening scenarios.

[0056] The operation module includes a code scanning unit for scanning the characteristic code and / or characteristic chip of the culture bottle, which is conducive to improving the informatization level of the cell culture equipment and facilitating traceability. Optionally, the code scanning unit is arranged in 1-3 groups and can be set as a scanning terminal, a camera, etc.

[0057] The operation module includes a centrifuge 1800 for centrifugal operation. The centrifuge 1800 can be an angle rotor centrifuge or a basket centrifuge with a temperature regulation function.

[0058] The operation module includes a microscope 1600 for observing cells and can also be used for cell status and counting.

[0059] The operation module includes an incubation unit 1022 for providing a preset incubation temperature. The incubation unit 1022 has multiple stations and can meet the constant temperature incubation requirements of culture bottles, culture dishes or well plates. The incubation temperature can be 37°C or other set temperatures.

[0060] It is worth noting that the robot arm 1011 can move consumables such as cell culture containers, 96-well plates, centrifuge tubes, etc. in space. The range of motion of the robot arm 1011 can cover more than 90% of the space of the working area 1001. For example, when the robot arm 1011 is set as a 6-axis robot arm 1011, the length of the working area 1001 is 1.5-2 times the length of the robot arm 1011. Optionally, multiple robot arms 1011 can also be provided.

[0061] like Figure 6-Figure 8As shown, the pipetting module is further provided with a wire supply assembly 4400, and the wire supply assembly 4400 includes a length-adjustable cable, one end of which is provided on the support seat 1015, and the free end is connected to the pipetting head 1016. By providing the wire supply assembly 4400 and connecting one end of the length-adjustable cable to the pipetting assembly 4100, when the pipetting assembly 4100 is separated from the support seat 1015 for pipetting, the length of the cable can be adjusted according to the distance between the pipetting assembly 4100 and the support seat 1015, thereby effectively reducing the entanglement between various leads of the pipetting assembly 4100 and other wires.

[0062] Specifically, the pipette head 1016 can be a component for sucking and discharging liquid. In one embodiment, the pipette head 1016 can be set as an independent pipette pump. In this case, the pipette pump needs wires for power supply and signal transmission; in other embodiments, the pipette head 1016 can also be a stainless steel pipette tip. In this case, the stainless steel pipette tip needs to be connected to a liquid tube for sucking and discharging liquid. Preferably, the pipette head 1016 is set as an ADP pipette.

[0063] Preferably, the pipetting head 1016 is detachably provided with a disposable nozzle, which is replaced after a pipetting to ensure the sterility of each pipetting and avoid contamination caused by multiple pipetting.

[0064] Furthermore, the cable can also retract or stretch when driven by other elastic components. Exemplarily, the cable supply assembly 4400 includes a cable reel 4410, one end of the cable is wound around the cable reel 4410, and the other end is fixedly connected to the pipetting assembly 4100. The pipetting assembly 4100 is provided with an electric wire or a liquid tube, and the electric wire or the liquid tube is fixedly connected to the cable, and the other end of the cable is connected to the pipetting assembly 4100. The cable reel 4410 can automatically reel in and unreel the cable and the electric wire or the liquid tube when the pipetting assembly 4100 moves, thereby reducing the entanglement between the leads. Optionally, the cable is set to a SUS stainless steel wire.

[0065] In other embodiments, the cable can also be a spiral wire or a spring wire that can be retracted under its own elastic force and can be stretched under the action of an external force. Exemplarily, the cable is set as a spring wire, and the spring wire is an electric wire to power the pipette head 1016; or the spring wire is a liquid tube connected to the pipette head 1016.

[0066] It is understandable that the longest retractable length of the cable is not less than the maximum distance between the pipetting component 4100 and the supporting seat 1015 .

[0067] One of the supporting seat 1015 and the pipetting body 4110 is provided with a male clamping head 4114, and the other is provided with a female clamping head 4111. The male clamping head 4114 can be clamped with the female clamping head 4111 to achieve the connection between the supporting seat 1015 and the pipetting body 4110. Specifically, the male clamping head 4114 is provided with a wedge-shaped protrusion, and the female clamping head 4111 is correspondingly provided with a clamping groove, and a bumping ball is provided inside the clamping groove. When the wedge-shaped protrusion is placed in the clamping groove, the bumping ball can limit the male clamping head 4114 in the clamping female head 4111. It can be understood that the pre-tightening force of the bumping ball is not less than the gravity of the pipetting assembly 4100 to ensure that the pipetting assembly 4100 can be stably placed in the supporting seat 1015. In other embodiments, the connection structure can also be set as a dovetail protrusion and dovetail groove that can be clamped with each other, or other elastic structures that can be clamped.

[0068] Preferably, the support seat 1015 is provided with a limiting platform, and the pipette body 4110 is provided with a limiting plate 4112. When the male clamping head 4114 is clamped with the female clamping head 4111, the limiting plate 4112 can be placed on the limiting platform. By providing the limiting platform and the limiting plate 4112 that cooperate with each other, it is used to assist in carrying the pipette assembly 4100, reduce the force between the male clamping head 4114 and the female clamping head 4111, and improve the reliability of placing the pipette assembly 4100 on the support seat 1015.

[0069] It is worth noting that the cell culture equipment is also provided with a control system, and the driving part and the detection part are both connected to the control system in communication. Optionally, the control system can adopt PID closed-loop control. The control system mainly consists of a memory, a sensor, a power source, a control component motion unit, etc. The memory can be considered as the central processing unit of the device, which can monitor the status of all positions of the device, the culture status of cell culture bottles, etc., including the culture time of the corresponding culture bottles in the cell culture equipment, and the cell culture status known by the microscope 1600 and other methods at the last moment. The sensor can include a temperature sensor, a flow sensor, a position optical coupler, etc. The power source can be a motor, a pump, etc., and the control components can be valves, position switches and other components.

[0070] The working process of the above-mentioned pipetting module is as follows: when the cell culture device receives an instruction that a cell culture bottle needs to be subcultured, the cell culture bottle that has completed digestion has been placed in the transfer position and is tilted at a certain angle to adapt to the pipetting head 1016; at the same time, the centrifuge tube that has been opened is placed in a relatively corresponding position. At this time, the mechanical arm 1011 drives the clamping component 4300 to approach the pipetting component 4100, and clamps the pipetting body 4110, and the machine moves horizontally or vertically to take the pipetting component 4100 out of the supporting seat 1015. Then the disposable suction nozzle of the pipetting head 1016 is replaced, and then the machine moves the pipetting component 4100 to the position of the cell culture bottle and absorbs the liquid. After a certain amount of liquid is absorbed, the machine moves the pipetting component 4100 to the nearby centrifuge tube again and spits out the liquid. When there is a large amount of liquid in the cell culture bottle or there are multiple liquids of the same type of cell culture bottles that need to be absorbed, the absorption can be completed by repeating the above actions. After the suction action is completed, the machine moves the pipetting component 4100 to the solid waste area, removes the disposable suction nozzle, and finally puts the pipetting component 4100 back to the supporting seat 1015.

[0071] It is worth noting that the movement of the pipette head 1016 can also be achieved by a multi-degree-of-freedom mechanism, such as an XYZ component. Furthermore, the process of pipetting in a centrifuge tube or a cell culture bottle can also use the pipette head 1016 to be fixed, and then the mechanical arm 1011 clamps the culture bottle or centrifuge tube and moves it to the pipette head 1016 to absorb and transfer the liquid. If multiple cell culture bottles or centrifuge tubes are operated in the same time period, the mechanical arm 1011 needs to continuously change hands to change different containers and move to the pipette head 1016 to absorb and receive liquid.

[0072] Further, the pipetting head 1016 of the pipetting module is arranged in the working area 1001 of the operation cabinet 1000, and the power components of the pipetting head 1016, such as pumps and valves, can be placed in the non-working area 1002. At the inlet end of the pump, if the pipeline is connected to a container containing disinfectant, the pipetting module can provide disinfectant for the container in the working area 1001, and when the inlet end of the pump is connected to a waste liquid bucket, the pipetting module can realize the waste liquid absorption function. Further, a sensor can be set at the inlet end of the pipetting head 1016 to monitor the presence or absence of liquid in the pipeline.

[0073] A well plate liquid transfer device can also be placed in the operation cabinet 1000. The well plate liquid transfer device can realize the functions of 96-well plate liquid transfer, 384-well plate liquid transfer, etc. In particular, the well plate liquid transfer device can be a single-channel liquid transfer pump or an 8-link liquid transfer pump, or a 96-channel or 384-channel liquid transfer assembly. The liquid transfer volume of a single channel is within 5ul-500ul, and 300ul can be selected preferably. The well plate liquid transfer device can be placed in an independent space with air filtration, or in the above-mentioned working area 1001. Preferably, the module is a 96-channel well plate liquid transfer assembly, and the 96-channel well plate liquid transfer assembly is installed in the working area 1001.

[0074] A cell monitoring device for cell fluorescence detection and activity detection is also arranged outside the non-working area 1002. The sample inlet end of the cell monitoring device is located in the working area 1001 or the inlet end can be connected to a transfer container, wherein the transfer container is arranged in the working area 1001, and the liquid pipeline between the inlet end and the cell culture device is provided with electromagnetic valves, pumps and other components, wherein the electromagnetic valve is used to monitor the cell monitoring device to close the liquid pipeline during non-working hours to prevent bacteria and other contaminants in the liquid pipeline from entering the working area 1001 of the cell culture device.

[0075] like Figure 9-11 As shown, the liquid storage module includes a storage box 5100, a plurality of liquid storage bottles 5200 and a detection unit 5300. A refrigeration unit 5110 is arranged in the storage box 5100. The cold end 5111 of the refrigeration unit 5110 is connected with the inner cavity 5101 of the storage box 5100 so that the inner cavity 5101 has a preset storage temperature; a plurality of liquid storage bottles 5200 are arranged in the inner cavity 5101, and the liquid storage bottles 5200 can be selectively connected with the main liquid pipe 5210; the detection module is arranged in the inner cavity 5101, and is arranged one-to-one with the liquid storage bottles 5200 to detect the remaining liquid in the liquid storage bottles 5200.

[0076] By providing a storage box 5100 with an inner cavity 5101, it is used to accommodate multiple liquid storage bottles 5200. Multiple liquid storage bottles 5200 can store one reagent or multiple reagents, which can increase the number and type of stored reagents. At the same time, the liquid storage bottle 5200 can be selectively connected to the main liquid pipe 5210 to ensure liquid supply; in addition, the detection unit 5300 is set in a one-to-one correspondence with the liquid storage bottle 5200, and can detect the remaining amount of reagent in the liquid storage bottle 5200 in real time, so that the reagent bottle and the main liquid pipe 5210 can be selectively connected or disconnected according to the remaining amount of reagent in the reagent bottle, thereby increasing the possibility of unattended operation of the liquid storage and supply device, thereby reducing manual regular replenishment of reagents, improving liquid supply efficiency, and reducing labor costs. By setting the cold end 5111 of the refrigeration unit 5110 to be connected to the inner cavity 5101, it is used for low-temperature storage environment.

[0077] Specifically, the cold end 5111 of the refrigeration unit 5110 is arranged on the side wall of the inner cavity 5101, and the refrigeration unit 5110 also includes a hot end 5112 and a cooling fan 5113, the hot end 5112 is connected to the cold end 5111, and is located outside the storage box 5100, that is, the refrigeration unit 5110 is penetrated through the side wall of the storage box 5100, and the refrigeration unit 5110 is installed by screws, which is convenient to install and easy to replace; the cooling fan 5113 placed outside the storage box 5100 can be used to dissipate heat from the hot end 5112, thereby ensuring the cooling effect of the cold end 5111. Optionally, the refrigeration unit 5110 is configured as a semiconductor refrigerator.

[0078] Furthermore, in order to prevent the temperature near the cold end 5111 of the refrigeration unit 5110 from being too low, which may cause the reagent in the liquid storage bottle 5200 near the cold end 5111 to freeze, a temperature-averaging fan 5120 is provided in the storage box 5100 to accelerate the gas flow in the inner cavity 5101 and to even out the temperature in the inner cavity 5101. Optionally, the temperature-averaging fan 5120 is configured as an axial flow fan.

[0079] Exemplarily, the preset storage temperature is set at 4° C.-8° C., and the preset use temperature is set at 30° C.-37° C. Optionally, the setting temperature of the cold end 5111 of the refrigeration unit 5110 is not less than 1.5° C. to prevent the temperature in the inner cavity 5101 from being too low and causing the reagent to freeze.

[0080] In this embodiment, the detection unit 5300 includes a weighing platform 5310, and the liquid storage bottle 5200 is arranged on the weighing platform 5310; optionally, the weighing platform 5310 includes a tray 5311 and a weight sensor 5312 arranged below the tray 5311, and the liquid storage bottle 5200 can be placed in the tray 5311. Preferably, a drainage through hole is also provided on the tray 5311, which is used to drain the condensed water in the tray 5311, so as to reduce the interference of the condensed water on the weight detection, and ensure the accuracy of the reagent residual detection.

[0081] In other embodiments, the detection unit 5300 includes a liquid level detection element to detect the liquid level of the liquid in the liquid storage bottle 5200, and the remaining amount of the reagent in the liquid storage bottle 5200 is determined by judging the liquid level. The liquid level can be detected by infrared or ultrasound, which is not specifically limited here.

[0082] In order to realize the selective communication between the multiple liquid storage bottles 5200 and the main liquid pipe 5210, the liquid storage and supply device further includes a control valve group 5500, which includes a plurality of switch valves 5510, and the plurality of switch valves 5510 are arranged one by one with the plurality of liquid storage bottles 5200. When the switch valve 5510 is placed in the open position, the liquid storage bottle 5200 and the main liquid pipe 5210 can be connected. It can be understood that the switch valve 5510 is connected to the detection unit 5300 of the corresponding liquid storage bottle 5200 in communication, so that the switch valve 5510 is selectively turned on and off according to the reagent residual amount detected by the detection unit 5300. In other embodiments, the control valve group 5500 can also be set as a multi-way valve, and the valve body of the multi-way valve is placed in different positions to realize the selective communication between the multiple liquid storage bottles 5200 and the main liquid pipe 5210.

[0083] Furthermore, the main liquid pipe 5210 is provided with a peristaltic pump 5211 , which is arranged upstream of the preheating unit 5400 and is used to extract the reagent in the liquid storage bottle 5200 when the liquid storage bottle 5200 is connected to the main liquid pipe 5210 .

[0084] Specifically, to ensure the storage temperature of the inner cavity 5101, the storage box 5100 is set to be double-layered, and the inner layer 5131 is set to be an insulation layer to reduce the low temperature of the inner cavity 5101 from spreading outward. At the same time, an insulation layer 6120 is set between the outer layer 5132 and the inner layer 5131 to further improve the insulation effect of the low temperature environment of the inner cavity 5101. Exemplarily, the insulation layer 6120 is set to be insulation cotton. Furthermore, the upper storage box is also provided with a door body to facilitate the placement of the liquid storage bottle 5200; and the door body is also provided with a heat preservation and heat insulation structure.

[0085] This embodiment also discloses a cell culture device, including the liquid storage and supply device as described in any of the above schemes. The cell culture device provided with the above liquid storage and supply device can store reagents in a low temperature storage environment, and the impact between multiple liquid storage bottles 5200 is small, and the reagents finally provided have a preset use temperature.

[0086] The use process of the above liquid supply module is as follows: when the liquid supply module is used, since the refrigeration unit 5110 is controlled by the PID control algorithm, the lowest temperature of the refrigeration unit 5110 will not be lower than the set threshold temperature (1.5°C); at the same time, since there is a temperature-averaging fan 5120 in the inner cavity 5101, it can be used to realize air circulation in the inner cavity 5101, so the temperature of the inner cavity 5101 is relatively uniform. When the detection unit 5300 of a certain liquid storage bottle 5200 detects that the residual amount of reagent in the corresponding liquid storage bottle 5200 is lower than the set threshold, the liquid storage bottle 5200 is disconnected from the main liquid pipe 5210 when the next test is added, and another liquid storage bottle 5200 is connected to the main liquid pipe 5210.

[0087] Exemplarily, cell culture equipment usually needs to place no less than two liquids for cell culture, such as trypsin liquid and cell culture medium liquid for culturing adherent cells. Taking into account the cell passaging and liquid replacement, the capacity of a single bottle of reagent is usually no less than 1L. The liquid storage and supply device can maintain unattended operation for at least 2 days. If unattended culture for a longer period of time such as 1 week is required, the corresponding reagent storage capacity needs to be increased, such as by placing 5-6 liquid storage bottles 5200 in the storage box 5100, and then the detection unit 5300 and the control valve group 5500 can be set accordingly.

[0088] like Figure 12-14 As shown, the liquid supply module includes a preheating unit 5400, the preheating unit 5400 includes a heating assembly 6300 and a consumable bag 6200, the consumable bag 6200 includes a bag body 6210, an outlet tube 6220 and an inlet tube 6230, one end of the outlet tube 6220 is connected to the bag body 6210, and the other end is connected to the pipetting module, one end of the inlet tube 6230 is connected to the bag body 6210, and the other end is connected to the liquid storage module, the end of the inlet tube 6230 away from the bag body 6210 is also connected to the air tube 6250, and the inlet tube 6230 can selectively connect to the liquid storage module and the air tube 6250.

[0089] A consumable bag 6200 is provided to hold the liquid to be preheated. A heating body 6100 is provided to heat the liquid to be preheated in the bag body 6210; specifically, the heating body 6100 is provided with a heating element 6310 and a heat-conducting block 6320, wherein the heating element 6310 is used to provide high temperature, and the heat-conducting block 6320 is provided between the heating element 6310 and the bag body 6210, for uniform temperature, and reducing the failure problem of the liquid to be preheated caused by excessive local temperature; and the heat-conducting block 6320 is provided with a placement groove, when the two heating bodies 6100 are buckled, the two placement grooves are buckled to form a placement space that can completely cover the bag body 6210, and the embracing heating can effectively ensure sufficient and uniform heating, and at the same time, the heating efficiency is higher. An inlet pipe 6230 and an outlet pipe 6220 are respectively provided at both ends of the bag body 6210 for the entry and exit of the liquid to be heated, wherein the inlet pipe 6230 is also connected to a liquid pipe 6240 and an air pipe 6250. When the liquid pipe 6240 is connected with the inlet pipe 6230, it is used to inject the liquid to be preheated into the bag body 6210. When the air pipe 6250 is connected with the inlet pipe 6230, it is used to drain the liquid in the bag body 6210, the inlet pipe 6230 and the outlet pipe 6220 to avoid waste.

[0090] Exemplarily, the heating element 6310 is configured as a PI heating film. The bag body 6210 has a thickness of 0.1 mm to 1 mm and a volume range of 1 ml to 20 ml, preferably a volume of 10 ml. The bag body 6210 can be a PP film, or a PP composite film, or can be provided with an aluminum layer. Preferably, a PP composite film with an aluminum thin layer is used.

[0091] Optionally, the inlet pipe 6230, the outlet pipe 6220 and the air pipe 6250 may be PVC pipes, silicone pipes, PU pipes, etc. The air pipe 6250 is provided with a filter, and when the air pipe 6250 and the inlet pipe 6230 are connected to drain the liquid, the air entering the consumable bag 6200 is filtered to avoid contamination of the liquid in the bag body 6210. A sensor may be provided on the outlet pipe 6220 to detect whether there is liquid passing through the outlet pipe 6220, and the sensor may be an ultrasonic flow sensor or an optical flow sensor. In addition, a peristaltic pump 5211 is provided on the inlet pipe 6230 to pump the liquid to be preheated into the bag body 6210.

[0092] Specifically, one end of the two heating bodies 6100 is rotatably connected to facilitate opening and closing operations. Exemplarily, the rotatable connection is achieved through a hinge 6130; a locking assembly 6110 is provided at the other end of the two heating bodies 6100 for locking the two heating bodies 6100 after the two heating bodies 6100 are buckled together.

[0093] Preferably, a heat preservation layer 6120 is provided on one side of the heating element 6310 away from the heat conductive block 6320. The heat preservation layer 6120 can wrap up the heat conductive block 6320, and only expose the side of the heat conductive block 6320 provided with the placement groove. After the consumable bag 6200 is installed in the heat conductive block 6320, the two heating bodies 6100 are buckled together, so that the bag body 6210 of the consumable bag 6200 is basically in a completely wrapped state for heating, and the heating effect is better. The structures of the two heating bodies 6100 are the same. In other embodiments, one of the heating bodies 6100 can also be set as the above structure, and the other heating body 6100 can be set as a metal plate with heat preservation function, a combination of heat preservation cotton or other plastic parts.

[0094] Furthermore, the heat conducting block 6320 is provided with a plurality of placement slots at intervals along its length direction, which are used to heat a plurality of consumable bags 6200 at the same time, thereby improving the ability of the liquid heating and supply device to supply more liquid at the same time, so as to adapt to more usage scenarios. It is understandable that the width of the placement slot is adapted to the width of the consumable bag 6200 after being filled with liquid. And the plurality of placement slots are independent of each other and do not affect each other.

[0095] Preferably, the preheating unit 5400 is also provided with a temperature detection member, which is used to detect the temperature of the placement space, and by detecting the temperature of the placement space, it is used to determine the temperature of the liquid in the consumable bag 6200. Optionally, the temperature detection member can be set in the heat conductive block 6320 of one of the heating bodies 6100. The temperature of the placement space can be set to 37°C.

[0096] In this embodiment, in order to realize that the inlet pipe 6230 can be selectively connected with the liquid pipe 6240 and the air pipe 6250, the consumable bag 6200 also includes a three-way valve, which is respectively connected to the inlet pipe 6230, the liquid pipe 6240 and the air pipe 6250, and the liquid pipe 6240 and the air pipe 6250 are opened and closed by controlling the position of the valve body of the three-way valve; or the consumable bag 6200 also includes two switch valves 5510, which are respectively arranged on the liquid pipe 6240 and the air pipe 6250, and the liquid pipe 6240 and the air pipe 6250 are opened and closed by respectively controlling the opening and closing of the switch valves 5510.

[0097] The volume of liquid in the inlet pipe 6230 is a, the volume of liquid in the bag body 6210 is b, the volume of liquid in the outlet pipe 6220 is c, and the required liquid volume is d; when d≤a+b+c, the liquid with a volume of d in the consumable bag 6200 is directly discharged through the outlet pipe 6220; when d>a+b+c, firstly, a liquid with a volume of 1.5 (dabc) is injected into the consumable bag 6200 through the inlet pipe 6230 and heated, and then the liquid with a volume of d in the consumable bag 6200 is discharged through the outlet pipe 6220.

[0098] The liquid supply method of the liquid supply module can effectively solve the problem of liquid waste caused by the volume of preheated liquid being larger than the actual required liquid volume in the prior art. Based on the structure of the liquid supply module, the required liquid volume is judged and compared with the total volume of the liquid in the bag body 6210, the inlet pipe 6230 and the outlet pipe 6220, and then the amount of liquid is controlled to be injected into the consumable bag 6200, so that when the liquid in the bag body 6210, the inlet pipe 6230 and the outlet pipe 6220 can meet the liquid supply once, the liquid is directly supplied; when the liquid in the bag body 6210, the inlet pipe 6230 and the outlet pipe 6220 cannot meet the liquid supply once, the liquid is first injected into the consumable bag 6200, and then the liquid is supplied. The amount of liquid injected is set with a certain safety margin to ensure the accuracy of liquid filling.

[0099] The use process of the above liquid supply module is as follows: the temperature of the placement space is set to a constant temperature such as 37°C, and the consumable bag 6200 is installed in the placement slot, and the heating body 6100 is buckled and locked. When the liquid injection instruction is received, the liquid pipe 6240 and the inlet pipe 6230 are connected, and the peristaltic pump 5211 starts to rotate, and the liquid in a bottle is pumped out, and flows into the bag body 6210 located in the placement space through the inlet pipe 6230, and then flows out to the outlet pipe 6220. When the control system determines that the current liquid filling test is the last test, it will fill the liquid according to the liquid supply method. When filling the liquid, the inlet pipe 6230 and the air pipe 6250 are connected, and the peristaltic pump 5211 rotates to allow air to flow into the inlet pipe 6230 after passing through the filter, and then the liquid in the consumable bag 6200 is filled into the corresponding container. If the remaining liquid in the consumable is less than or equal to the required filling amount, the liquid pipe 6240 and the inlet pipe 6230 are first connected, and the peristaltic pump 5211 rotates to extract a certain amount of liquid. After the extraction is completed, the inlet pipe 6230 and the air pipe 6250 are connected, and the peristaltic pump 5211 keeps rotating, so that air flows into the inlet pipe 6230 through the filter. After the liquid filling is completed, all the remaining liquid can be discharged into the waste liquid area.

[0100] Generally speaking, only one type of cell will be cultured in the same batch in a cell workstation, so the growth rate and growth mode of cells in certain specific culture containers are known in advance. The parameters known in advance include: the culture environment temperature required for primary cell culture, carbon dioxide concentration, the time it takes for primary cells to grow to be passaged, the time it takes for cells to be cultured from one pass to the second pass, the time it takes for cells to be replaced with liquid, the amount of reagents added, etc. The control system can estimate in advance the number of times that passaging and liquid changes are required on the day, and thus calculate the amount of various liquids that need to be added on the day.

[0101] An operating table is provided in the operating cabinet 1000, and the operating table is used to separate the working area 1001 from the non-working area 1002. A waste opening 1900 and an isolation door are provided on the operating table, and the isolation door can selectively block the waste opening 1900 to selectively connect the waste area with the working area 1001. Furthermore, the waste area is also provided with a waste carrier 1701, a waste switch door and a sliding assembly, and the waste carrier 1701 is provided on the sliding assembly, and the sliding assembly is used to move the waste carrier 1701 out of the waste switch door.

[0102] The waste port 1900 is used to discharge solid waste and liquid waste. A waste carrier 1701 for receiving waste is stored below the waste port 1900 (the waste carrier 1701 is in the non-working area 1002). Optionally, the waste carrier 1701 is placed on a retractable drawer, and the drawer can be further equipped with an electric device to realize the extension and recovery movement of the drawer.

[0103] The waste port 1900 is provided with a waste switch door, which can be opened by the drive of an electric device. Furthermore, the waste switch door can be rotated to open or moved. A sealing strip is provided at the matching position on the other side of the waste switch door to prevent the air of the waste carrier 1701 from flowing into the working area 1001. In particular, when the door is swung open, the direction of the waste switch door is the direction close to the robot arm 1011. The waste port 1900 is provided as a trumpet-shaped contraction port, and the waste can flow out of the contraction port to the waste carrier 1701 under the action of gravity. In particular, a switch valve for controlling the on-off of the pipeline is provided between the waste carrier 1701 and the trumpet-shaped contraction port. Liquid waste and solid waste can be discarded in the same waste carrier 1701 or in different waste carriers 1701. Furthermore, a sensor is provided below the waste carrier 1701 to detect the full load of materials in the waste carrier 1701. The signal can be used to mobilize the handling robot to replace the waste carrier 1701, or to warn of the full load of waste. The sensor can be a weighing sensor. The waste carrier 1701 can be set as a waste barrel.

[0104] The cell culture equipment also includes a handling robot to carry out transportation between the culture cabinet 2000, the operation cabinet 1000 and the consumable cabinet 3000. The handling robot has a moving part and a moving arm part. The moving arm can be a 6-axis robot 1011 or a collaborative robot commonly seen on the market. The handling robot can cooperate with the control instructions of the cell culture equipment or the instructions of the operator to replenish the consumable materials or carry and update the waste barrel. The handling robot can also dock the laboratory's biosafety transfer window with the designated docking station on the cell culture equipment. Further, the docking station of the cell culture equipment is not less than the sum of the number of the feeding docking mechanism of the consumable cabinet 3000, the number of the waste barrel station, and the operating station of the operation cabinet 1000. For example, the number of stations of the feeding docking mechanism of the consumable cabinet 3000 is 2, the number of stations of the waste barrel is 1, and the number of stations of the safety transfer window is 1, then the number of stations that the mobile robot can dock is not less than 4.

[0105] Exemplarily, the above-mentioned cell culture device can perform the following operations:

[0106] Cell culture: During operation, when the cell culture flask in the culture cabinet 2000 needs to be replaced with liquid, the bottle-taking mechanism of the culture cabinet 2000 will take the cell culture flask out of the box in the culture cabinet 2000. At this time, the sealed door located on the side of the culture cabinet 2000 close to the operating cabinet 1000 needs to be opened, and then the cell culture flask is sent to the first window 1300 of the operating cabinet 1000 that is connected to the incubator. A code scanning unit is arranged on the side of the first window 1300. When the cell culture flask is sent out, the code scanning unit can identify the barcode or RFID chip on the cell culture flask to record the status of the cell culture flask. After the culture flask is sent out, the multi-degree-of-freedom motion mechanism located in the operating cabinet 1000, such as the 6-axis robot arm 1011, will be driven to the position of the cell culture flask to clamp the culture flask. The end of the 6-axis robot arm 1011 is equipped with an actuator. After clamping the culture flask, the bottle is transferred to the bottom of the opening unit 1500 to open the lid of the culture flask. After the opening is completed, the cell culture bottle cap will be retained in the clamp of the opening unit 1500. At this time, the robot arm 1011 moves the culture bottle to the dumping position of the liquid waste, and dumps the culture bottle in the culture bottle at this time. At the same time, the solenoid valve and pump under the waste port 1900 are started to absorb the waste liquid. The working area 1001 is also provided with a liquid filling module. Then the robot arm 1011 moves the cell culture bottle to the bottom of the liquid filling module to add fresh culture medium to the cells. At this time, the pump and solenoid valve on the pipeline corresponding to the liquid filling module work to pump out the refrigerated culture medium in the storage box 5100 and drive the liquid to flow through the preheating unit 5400, and then the corresponding volume of culture medium is added to the cell culture bottle. After the action is completed, the robot arm 1011 moves the culture bottle to the bottom of the corresponding opening unit 1500, closes the cover of the culture bottle, and then returns it to the designated position of the first window 1300 where the operation cabinet 1000 and the incubator are docked. The culture bottle is sent back to the incubator 2000 to continue incubation and growth.

[0107] Subculture work: When the cell culture bottle reaches the corresponding replacement time, the incubator will send the corresponding cell culture bottle from the incubator to the specified position of the first window 1300 where the operating cabinet 1000 is connected to the incubator. At this time, the barcode scanner or RFID sensor of the scanning unit will read the information of the culture bottle, and then the robot arm 1011 will move the culture bottle to the observation platform of the microscope 1600. The microscope 1600 reads the cell status in the culture bottle. When the software determines that the cells in the cell culture bottle meet the conditions for subculture, the robot arm 1011 will take the culture bottle out to the opening unit 1500 and open the cover. Then, the culture bottle is transferred to the waste port 1900 to dump the waste liquid. At the same time, the pump and solenoid valve for extracting the waste liquid are opened to absorb the waste liquid. Then the robot arm 1011 moves the culture bottle to the corresponding filling port of the liquid filling module, adds cell culture fluid to clean the residual liquid of the cells, and pours out the liquid after cleaning again. The number of cleanings can be 1 time or more, which is determined according to actual needs. Then the robot arm 1011 moves the culture bottle to the bottom of the opening unit 1500, closes the bottle cap, and then places the culture bottle in the incubation unit 1022 located in the working area 1001 of the operating cabinet 1000 for incubation.

[0108] Observation of cell growth status: Then the robot 1011 takes out another cell culture bottle from the culture cabinet 2000, repeats the above steps, and then places the culture bottle in the second position of the incubation unit 1022. It should be noted that during the observation stage, when the microscope 1600 reads that the cell growth in the cell culture bottle does not meet the passage conditions, the cell culture bottle will be directly returned to the culture cabinet 2000 to continue incubation and growth. The incubation unit 1022 has no less than 2, preferably 4 incubation positions, namely #1, #2, #3, and #4.

[0109] Digestion and termination of digestion: Robotic arm 1011 takes out the cell culture bottle on the #1 incubation position, opens the cover, pours the liquid, and then goes to the liquid filling device to add digestive enzymes. After the digestive enzymes are added, the cell culture bottle closes the cover and is returned to the #1 incubation area. Then, the cell culture bottle on the #2 incubation position is taken out to open the cover, pour the liquid, and similarly add digestive enzymes. After the liquid is added, it is returned to the #2 incubation area. When the digestion time is up, robot arm 1011 will open the covers of the cell culture bottles in sequence, move to the corresponding position of the liquid filling device to terminate the digestion liquid filling, close the covers, shake the culture bottles, and then return the culture bottles to the corresponding incubation areas.

[0110] Centrifuge tube preparation: At this time, the robot arm 1011 can go to the centrifuge tube consumable position to clamp a new centrifuge tube. After clamping the centrifuge tube, it is transferred to the bottom of the opening unit 1500 to open the centrifuge tube cover, and then the centrifuge tube is transferred to the liquid transfer position of the centrifuge tube. Considering that multiple bottles of cells can be passaged, the centrifuge tubes can take the same number as the cell culture bottles that need to be passaged at the same time and place them in the liquid transfer position.

[0111] Transfer of cell culture flasks: Place the cell culture flasks of the incubation unit 1022 at the position for changing hands for the flasks, change the way the robot arm 1011 holds the cell culture flasks, open the flasks, and place them near the liquid aspiration position of the pipette head 1016. It should be noted that the liquid aspiration position of the cell culture flasks has a certain inclination angle, so that when the pipette head 1016 moves vertically into the flask, the end of the pipette head 1016 can reach the lowest point in the flask. If there are multiple flasks, these operations can be performed one after another.

[0112] Liquid transfer: The robot arm 1011 clamps the liquid transfer head 1016 and makes the robot arm 1011 grab the liquid transfer head 1016, and then transfers the liquid in the culture bottle to the centrifuge tube. After the liquid transfer is completed, the liquid transfer head 1016 is hung back to the original position (supporting seat 1015).

[0113] Centrifuge tube centrifugation: Close the caps of the centrifuge tubes one by one and place them in the corresponding hole positions of the centrifuge 1800. It should be noted that if the number of centrifuge tubes is odd, the centrifuge 1800 needs to be balanced. During the waiting time for centrifugation, the robot 1011 can move the old cell culture bottle to the bottom of the filling port of the disinfectant filling device 1026 at a time, and add liquids such as alcohol or 84 disinfectant, and then close the cap of the culture bottle and discard it into the waste bucket. Furthermore, an old cell culture bottle can be left in the transfer position to hold the supernatant of the centrifuge tube after centrifugation.

[0114] Cell resuspension and liquid transfer after centrifugation: After centrifugation, the robot arm 1011 clamps the centrifuge tube, opens the cover, and then puts it back to the transfer position 1027 of the centrifuge tube, and then moves the upper liquid after centrifugation to the old culture bottle, and then hangs the pipette head 1016 back, and then adds disinfectant to the old culture bottle and closes the cover and discards it. After that, clamp the centrifuge tube and move it to the liquid filling device to add the corresponding culture liquid for liquid resuspension, and finally put the centrifuge tube back to the transfer position.

[0115] Preparation of new culture bottles: Pick up new culture bottles in the culture bottle consumables area and identify the information. Then open the lid of the culture bottle and move the culture bottle to the pipetting and filling device to add culture medium. Then place it on the pipetting transfer position 1027. The number of new culture bottles is picked up according to the actual needs.

[0116] Resuspended cell mixing and packaging: The robot arm 1011 clamps the pipette head 1016 to absorb the mixed liquid in the centrifuge tube, and transfers all the liquid to the same centrifuge tube, then blows the liquid several times, and then absorbs a certain amount of the mixed liquid and transfers it to a new culture bottle. Furthermore, when transferring a new cell mixture, a new pipette head 1016 can be replaced.

[0117] The cells after passage are returned to the culture bottle: the culture bottle that has been packed with cells is covered and transferred to the transfer position of the first window 1300 of the operation cabinet 1000 and the incubator, and then the cell culture bottle is sent back to the incubator 2000.

[0118] Drug transfer and culture: When the cells grow to a certain extent, the cell mixture in the culture bottle needs to be transferred to the well plate. At this time, the single-well plate is first taken out from the consumable cabinet 3000 and placed in the position where the culture bottle is changed. Then the digested and terminated cell solution is poured onto the single-well plate. Then the single-well plate is transferred to the working position of the pipetting module, and the 96-channel pipetting device 1018 is driven to aspirate. After the aspiration is completed, the single-well plate is transferred to the disinfectant filling port, a certain amount of disinfectant is added, and then the entire single-well plate is discarded. Then the 96-well plate with the drug solution is transferred from the consumable cabinet 3000 or transferred to the cell culture bottle handover position of the operation cabinet 1000 by the handling robot, and the mechanical arm 1011 clamps the 96-well plate and moves the information identification area 1019, reads and records the information of the well plate, and then moves to the bottom of the electric suction cup cover opener 1510 to open the lid of the 96-well plate, and then the well plate is placed at the transfer position for the 96-well plate, and then the cell solution is added to the well plate with the drug, and the liquid can be further blown slightly. Finally, the well plate is clamped and moved to the electric suction cup cover opener 1510, the lid is taken back, and then the well plate is transferred to the culture cabinet 2000. Further, when drug screening and virus culture are performed, the culture conditions in the culture cabinet 2000 can be set according to the actual situation. At this time, the mixed liquid transfer in the cell culture bottle in the box of the original culture cabinet 2000 needs to be transferred to the corresponding well plate.

[0119] Consumables replenishment: When the remaining consumables in the consumable cabinet 3000 are insufficient, a reminder signal can be sent to remind the staff to replenish the consumables. Furthermore, the consumables can be replenished manually or by a transport robot, which takes the consumables out from the designated laboratory transfer window and then transfers them to the consumable transfer position designated in the consumable cabinet 3000 to complete the consumables replenishment.

[0120] Waste barrel transportation: When the capacity of the waste barrel is insufficient, the equipment can send a prompt signal to remind the staff to change the waste barrel. Furthermore, the waste barrel can be replenished manually or by a handling robot. A new waste barrel can be taken out from a designated laboratory transfer window or other location and then replaced in the position of the waste barrel on the equipment to complete the replacement of the waste barrel.

[0121] The above contents are only preferred embodiments of the present invention. For ordinary technicians in this field, according to the concept of the present invention, there will be changes in the specific implementation methods and application scopes. The content of this specification should not be understood as limiting the present invention.

Claims

1. A cell culture device, characterized in that include: An operating cabinet (1000), wherein a working area (1001) and a non-working area (1002) are arranged in the operating cabinet (1000), wherein the working area (1001) is provided with a mechanical arm (1011), a liquid supply module, a liquid transfer module and at least two operating modules, wherein the non-working area (1002) is provided with a liquid storage module and a waste area, wherein the liquid storage module is used to store liquid at a preset storage temperature, and the liquid supply module is connected to the liquid storage module and is used to heat a preset amount of liquid to a preset use temperature; wherein the working area (1001) is provided with a mechanical arm (1011), a liquid supply module, a liquid transfer module and at least two operating modules, wherein the non-working area (1002) is provided with a liquid storage module and a waste area, wherein the liquid storage module is used to store liquid at a preset storage temperature, and wherein the liquid supply module is connected to the liquid storage module and is used to heat a preset amount of liquid to a preset use temperature; 001) domain is also provided with a bearing seat (1015), the pipetting module (1013) is provided with a pipetting head (1016), the pipetting head (1016) is detachably provided on the bearing seat (1015), the mechanical arm (1011) can selectively clamp and drive the pipetting head (1016) to perform pipetting; the mechanical arm (1011) can move and operate between the pipetting module (1013) and two or more operation modules; the waste area is selectively connected to the working area (1001); The culture cabinet (2000) and the consumable cabinet (3000) are respectively arranged on both sides of the operation cabinet (1000) and can be selectively connected to the operation cabinet (1000); the culture cabinet (2000) is used for cell culture, and the consumable cabinet (3000) is used for storing consumables.

2. The cell culture device according to claim 1, characterized in that: The pipetting module (1013) is also provided with a wire supply assembly (4400), and the wire supply assembly (4400) includes a cable with adjustable length, one end of the cable is arranged on the supporting seat (1015), and the free end is connected to the pipetting head (1016).

3. The cell culture device according to claim 1, characterized in that: The liquid storage module comprises a storage box (5100), a plurality of liquid storage bottles (5200) and a detection unit (5300); a refrigeration unit (5110) is arranged in the storage box (5100); a cold end (5111) of the refrigeration unit (5110) is communicated with an inner cavity (5101) of the storage box (5100) so that the inner cavity (5101) has a preset storage temperature; a plurality of liquid storage bottles (5200) are arranged in the inner cavity (5101), and the liquid storage bottles (5200) can be selectively connected to the main liquid pipe (5210); a detection module is arranged in the inner cavity (5101), and is arranged one-to-one with the liquid storage bottles (5200) so as to detect the residual amount of liquid in the liquid storage bottles (5200).

4. The cell culture device according to claim 1, characterized in that: The liquid supply module includes a preheating unit (5400), and the preheating unit (5400) includes a heating component (6300) and a consumable bag (6200). The consumable bag (6200) includes a bag body (6210), an outlet pipe (6220) and an inlet pipe (6230). One end of the outlet pipe (6220) is connected to the bag body (6210), and the other end is connected to the pipetting module (1013). One end of the inlet pipe (6230) is connected to the bag body (6210), and the other end is connected to the liquid storage module. The end of the inlet pipe (6230) away from the bag body (6210) is also connected to the air pipe (6250), and the inlet pipe (6230) can selectively connect to the liquid storage module and the air pipe (6250).

5. The cell culture device according to claim 1, characterized in that: An operating table is arranged in the operating cabinet (1000), and the operating table is used to separate the working area (1001) and the non-working area (1002). A waste port (1900) and an isolation door are arranged on the operating table, and the isolation door can selectively block the waste port (1900) to selectively connect the waste area and the working area (1001).

6. The cell culture device according to claim 5, characterized in that: The waste area is also provided with a waste carrier (1701), a waste switch door and a sliding assembly. The waste carrier (1701) is arranged on the sliding assembly, and the sliding assembly is used to move the waste carrier (1701) out of the waste switch door.

7. The cell culture device according to claim 1, characterized in that: The operation module comprises a cover opening unit (1500) for opening the centrifuge tube and / or the culture bottle; and / or the operation module comprises an incubation unit (1022) for providing a preset incubation temperature.

8. The cell culture device according to claim 1, characterized in that: The operation module further comprises a filtering unit (1017) for filtering and circulating the air in the working area (1001); and / or the operation module comprises a scanning unit for scanning the characteristic code and / or characteristic chip of the culture bottle and / or centrifuge tube.

9. The cell culture device according to claim 1, characterized in that: The culture cabinet (2000) is provided with a sterilization module for providing hydrogen peroxide fumigation operation.

10. The cell culture device according to any one of claims 1 to 9, characterized in that: The cell culture device further comprises a transport robot for carrying out transport between the culture cabinet (2000), the operation cabinet (1000) and the consumables cabinet (3000).

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