CN117866763A - Automatic change cell culture studio and equipment - Google Patents

Abstract

The invention relates to the field of cell culture equipment, in particular to an automatic cell culture working chamber, which comprises a lower working area, a middle working area and an upper working area; the lower working area is provided with a coding assembly, a microscope assembly, a centrifugal machine and an electric assembly, and the middle working area is provided with a sterilizing mechanism, a rotary table for storing culture bottles, a vial sample adding assembly, a consumable rack, a cell counter, a refrigerator for storing cells, an inner manipulator and a vial rack; the upper working area is provided with a triaxial mechanical arm and a hepa system, and the triaxial mechanical arm is used for being matched with the middle working area. The device has small occupied area and compact structure, and simplifies the automatic execution of cell culture.

Description

Automatic change cell culture studio and equipment

Technical Field

The invention relates to the field of cell culture equipment, in particular to an automatic cell culture working chamber and equipment.

Background

The existing market lacks automatic cell culture equipment or instruments, and has more flows of cell culture, cell harvest, data recording, consumable sterilization and the like, and the existing cell culture equipment has incomplete functions.

Because the cell culture process needs a complicated preparation process, a plurality of instruments are needed to be matched with each other, the instruments occupy a large space, the space distribution of the design of a plurality of instruments is unreasonable, so that the equipment is huge and complex, and the execution of the process is inconvenient.

Disclosure of Invention

In order to solve the problem of overlarge volume of the cell culture equipment, the invention provides an automatic cell culture equipment which has small occupied area and compact structure, simplifies the automatic execution of cell culture and adopts the following technical scheme:

an automated cell culture working chamber comprising a lower working area, a middle working area and an upper working area;

the lower working area is provided with a coding assembly, a microscope assembly, a centrifugal machine and an electric assembly,

the middle working area is provided with a sterilizing mechanism, a rotary table for storing culture bottles, a vial sample adding assembly, a consumable rack, a cell counter, a refrigerator for storing cells, an inner manipulator and a vial rack;

the upper working area is provided with a triaxial mechanical arm and a hepa system, and the triaxial mechanical arm is used for being matched with the middle working area.

The automatic cell culture working chamber is characterized in that the coding assembly, the microscope assembly and the centrifugal machine are sequentially arranged in front of the lower working area from left to right;

the inlet of the centrifugal machine is communicated with the middle working area, and the rotary table, the sterilizing mechanism and the inlet of the centrifugal machine are sequentially arranged in front of the middle working area from left to right; the refrigerator, the vial sample adding assembly and the inner manipulator are sequentially arranged in the middle position of the middle working area from left to right; the cell counter, the consumable rack and the vial rack are sequentially arranged at the rear position of the middle working area from left to right;

the triaxial mechanical arm is matched with the left side area of the middle working area.

The automatic cell culture working chamber is divided into a left working chamber and a right working chamber;

the left working chamber is internally provided with a horizontal left working table plate to form a separation of a lower working area and a middle working area, the code printing component and the microscope component are positioned in front of the lower working area below the left working table plate, the rotary table is positioned in front of the left working table plate, the refrigerator and the vial sample adding component are positioned in the middle of the left working table plate, the cell counter and the consumable rack are positioned in the rear of the left working table plate, and the triaxial mechanical arm is positioned in the upper working area of the left working chamber;

the right working chamber is internally provided with a horizontal right working table plate to form a separation of a lower working area and a middle working area, the centrifugal machine is positioned on the right side of the front position of the lower working area below the right working table plate, the inlet of the centrifugal machine is communicated with the right working table plate, the sterilizing mechanism and the inlet of the centrifugal machine are positioned on the front position of the right working table plate, the inner manipulator is positioned in the middle position of the right working table plate, and the bottle rack is positioned at the rear position of the right working table plate.

The automatic cell culture working chamber comprises a laser marking machine, a protective cover I and a conveying mechanism, wherein the laser marking machine comprises a machine base and a marking head, the machine base is positioned in the protective cover I, the conveying mechanism is arranged on the machine base, an opening for passing laser of the marking head is formed in the top of the protective cover I, and an opening for the telescopic in-out of the conveying mechanism is formed in the front side of the protective cover I;

the top of the shield I is provided with a smoke pouring pipe and a fan, and the fan is aligned to the smoke pouring pipe.

The automatic cell culture working chamber is characterized in that the rotary table is provided with a plurality of bottle clamp assemblies, each bottle clamp assembly comprises an openable clamp, an electric clamp groove for controlling the clamp, a bracket and a steering engine, the lower end of the steering engine is fixedly arranged on a rotary base of the rotary table, the bracket is rotatably arranged at the upper end of the steering engine, the electric clamp groove is arranged on the bracket, the clamp is correspondingly arranged at the output end of the electric clamp groove,

the automatic cell culture working chamber is characterized in that a heating plate is arranged on the clamp.

The automatic cell culture working chamber comprises a groove frame for storing culture medium bottles, a placing table for storing covers and a plurality of clamping jaws for clamping the bottles.

Above-mentioned automatic change cell culture studio, disinfection mechanism includes keeps apart storehouse, automatically controlled apron II that opens and shuts, is used for placing carrier, fan and the shower nozzle of blake bottle, keeps apart the front side in storehouse and is the opening, and apron II is located the top in keeping apart the storehouse, and the carrier is installed in keeping apart the storehouse, and fan and two shower nozzles are established at the inside rear side in keeping apart the storehouse and just to the carrier.

Above-mentioned automatic change cell culture studio, the rear bit of middle part workspace still is equipped with the weighing machine that is used for weighing the culture medium bottle, the middle position of middle part workspace still is equipped with the waste liquid bucket, the waste liquid bucket is located the right side of interior manipulator.

The invention also relates to an automatic cell culture device which comprises a working chamber, a plurality of incubators with empty bottle racks and an external manipulator, wherein the working chamber is the automatic cell culture working chamber.

Above-mentioned automatic change cell culture equipment, outer manipulator execution end includes tray anchor clamps, sucking disc, guard shield II and mounting panel, through the mounting panel will carry out the end and be connected with outer manipulator main part, be equipped with terminal holder, controller and air pump in the guard shield II.

The invention has the beneficial effects that:

firstly, the composition and arrangement of each component in a working chamber are optimized, the whole machine has compact structure and small occupied area, the structural change also simplifies the automatic execution logic, the complex procedures of disinfection, cultivation, analysis, harvest and the like are rapidly and accurately completed, and the efficiency of cell culture is greatly improved.

Secondly, the code printing component adopts laser, is not easy to wear, and compared with code pasting, the code printing component has the advantages that the problems that the code cannot be scanned due to blurring caused by factors such as disinfection and the like are solved.

Thirdly, functions such as application of sample subassembly, consumptive material frame integrate, are applicable to the vial equipment of different specifications, further simplify the control.

Drawings

FIGS. 1 and 2 are perspective views showing the overall structure of the apparatus according to the embodiment of the present invention;

FIG. 3 is a schematic top view of the overall structure of the apparatus according to the embodiment of the present invention;

FIG. 4 is a schematic top view of a working chamber according to an embodiment of the present invention;

FIG. 5 is a perspective view of a left working chamber according to an embodiment of the present invention;

fig. 6 is an internal top view of the left working chamber (the inner manipulator 25 omits part of the structure) according to the embodiment of the present invention;

FIGS. 7 and 8 are schematic diagrams of a frame of a left working chamber according to an embodiment of the present invention;

FIG. 9 is a schematic diagram of a coding module according to an embodiment of the present invention;

FIG. 10 is a schematic view of a left table assembly according to an embodiment of the invention;

FIG. 11 is a schematic view of a bottle clip assembly according to an embodiment of the present invention;

FIG. 12 is a schematic view of a Z-axis assembly of a three-axis mechanical arm according to an embodiment of the present invention;

FIG. 13 is a perspective view of a right working chamber according to an embodiment of the present invention;

FIG. 14 is a top view of a right working chamber according to an embodiment of the present invention;

FIG. 15 is a perspective view of a right working chamber according to another embodiment of the present invention;

FIG. 16 is a schematic perspective view of a sterilization mechanism according to an embodiment of the present invention;

FIG. 17 is a schematic view showing the internal structure of a sterilizing mechanism according to an embodiment of the present invention;

FIG. 18 is a schematic view of an outer manipulator execution end structure according to an embodiment of the present invention;

fig. 19 is a schematic view of an internal structure of an external manipulator execution end according to an embodiment of the present invention.

In the figure: left working chamber 1, left housing 11, coding module 12, microscope module 13, left working table 14, turntable 15, tank rack 16, placement table 17, three-axis mechanical arm 18, consumable rack 19, cell counter 111, refrigerator 112, dust detection module 113, and clamping jaw 114;

conveying mechanism 121, shield I122, base 123, marking head 124, inverted smoke tube 125, fan 126;

the bottle cap holder comprises a clamp 151, a bottle cap seat 152, an electric clamping groove 153, a bracket 154, a steering engine 155 and a heating plate 156;

suction cup I181, uncapping clamping jaw 182, large plug connector 183 and small plug connector 184;

the right working chamber 2, the right outer cover 21, the sterilizing mechanism 22, the centrifugal machine 23, the right working platen 24, the inner manipulator 25, the weighing device 26, the bottle rack 27, the waste liquid barrel 28, the bin gate 211, the cover plate I212 and the code scanner 213;

isolation bin 221, cover plate II 222, carrier 223, fan 224, and spray head 225;

empty bottle rack 3, incubator 4, outer manipulator 5, tray clamp 51, suction cup II 52, shield II 53, end gripper 54, controller 55, air pump 56, mounting plate 57, hepa system 6, culture bottle 10, culture medium bottle 20, small centrifuge tube 30, large centrifuge tube 40.

Detailed Description

The technical scheme of the invention is described in detail below with reference to the accompanying drawings.

The embodiment is an automatic cell culture device, each component layout is reasonable, and the whole machine size is compact. As shown in fig. 1 to 3, the apparatus mainly comprises a working chamber, an empty bottle holder 3 for storing culture bottles 10, three incubators 4, and an external manipulator 5 for holding transfer culture bottles 10, and a hepa system 6 is installed above the working chamber for ensuring cleanliness. The culture flask 10 completes the earlier processes of coding, sterilizing, filling cells, culture medium and the like in a working chamber, the external manipulator 5 places the culture flask 10 into the culture box 4 for culture, microscopic observation is kept during the culture period, and the culture flask 10 enters the working chamber again for harvesting processes (cell cleaning, pancreatin digestion, neutralization, centrifugation and the like) after the culture is completed.

The empty bottle rack 3 and the incubator 4 of the present embodiment are substantially identical to the structure disclosed in the previous generation device CN115895894a, and will not be described in detail herein.

The working chamber is structurally designed into a lower working area, a middle working area and an upper working area. The lower working area is mainly used for installing electrical components, coding components 12, microscope components 13, centrifuges 23, etc. The middle working area is provided with a workbench, and a code scanner 213, a sterilizing mechanism 22 for sterilizing, a turntable 15 for storing culture bottles 10, a groove frame 16 for storing culture medium bottles 20, clamping jaws for clamping centrifuge tubes, a consumable frame 19 for storing consumables such as tip tubes, a cell counter 111, a refrigerator 112 for storing cells, an inner manipulator 25, a weighing device 26 for weighing the culture medium bottles 20, a bottle frame 27 for storing centrifuge tubes and the like, a waste liquid barrel 28 for storing waste culture liquid and the like are arranged on the workbench. The three-axis mechanical arm 18, the monitoring camera, the hepa system 6 and the like are arranged at the top, namely the upper working area, and the three-axis mechanical arm 18 can not only finish taking, putting and uncovering containers such as centrifuge tubes, cell tubes and culture medium bottles 20, but also take tip tubes to finish harvesting cells.

In this embodiment, the working chambers are divided into a left working chamber 1 and a right working chamber 2, and the left working chamber and the right working chamber are communicated, so that the inner manipulator 25 is convenient to operate, the positions of the devices in the working area are reasonably arranged, and meanwhile, the process steps are optimized, so that the efficiency of the earlier process and the subsequent harvest of the cells is greatly improved. As shown in FIG. 3, the working chamber has a lateral length of about 2 meters and a vertical length of about 1.6 meters, is compact in structure and small in occupied space, and the total occupied area of the working chamber, the empty bottle rack 3, the incubator 4 and the like is about 15 square meters, so that the occupied area of the working chamber is obviously reduced compared with that of the previous generation of products.

Left working chamber 1

As shown in fig. 5 to 8, the left working chamber 1 is of a frame structure as a whole, a left outer cover 11 is externally mounted, and switch doors for observation, debugging, overhaul and the like are correspondingly arranged on the front, rear, left and three surfaces of the left outer cover 11.

(1) Lower working area of left working chamber 1

The lower working area of the left working room 1 is used for installing a coding assembly 12, a microscope assembly 13, a distribution box and the like, and the coding assembly 12 and the microscope assembly 13 are both provided with conveying mechanisms for conveying the culture bottles 10 into corresponding areas to realize coding or microscopic observation. The coding and microscopic observation steps are all conveying mechanisms for the outer manipulator 5 to grasp the culture flask 10 and put it in the corresponding step, and for convenience of execution, as shown in fig. 5, the coding assembly 12 and the conveying mechanism for the microscope assembly 13 are both disposed on the front side of the left working chamber 1.

The culture flask 10 is numbered and coded prior to use. As shown in FIG. 9, the coding assembly 12 selects a laser marker to code the culture flask 10, and comprises a telescopic conveying mechanism 121, a shield I122, a machine base 123, a marking head 124, a smoke pouring pipe 125 and a fan 126. The frame 123, the stand and the marking head 124 that disposes the laser instrument are the main component of laser marking machine, and conveying mechanism 121's fixed plate is on the frame 123, and guard shield I122 is used for covering frame 123, and the top of guard shield I122 is equipped with the opening that is used for marking head 124 laser to pass through, and the front side of guard shield I122 sets up the opening that is used for conveying mechanism 121 to stretch out and draw back in and go out corresponding to the front side of left working chamber 1. The laser coding is to etch the surface of the culture flask 10, and compared with the prior code pasting, the laser coding has no problems of fuzzy code number, abrasion and the like caused by the procedures of alcohol disinfection and the like. In order to reduce smoke dust, a smoke pouring pipe 125 and a fan 126 for pumping out marking smoke dust are further arranged at the top of the protecting cover I122, so that the smoke dust in the protecting cover I122 is discharged in time, and pollution to the culture flask 10 is reduced.

In the process of culturing cells in the incubator 4, the external manipulator 5 takes out the culture flask 10 in the incubator 4, places the culture flask 10 at the microscope assembly 13 for microscopic observation, records the growth condition of the cells, and then the external manipulator 5 returns the culture flask 10 to the incubator 4 for continuous culture. After the cultivation is completed, the external manipulator 5 sends the culture flask 10 into the working chamber for the harvesting process. The microscope assembly 13 of the present embodiment is substantially identical to the structure and function disclosed in the previous generation device CN115895894a, and will not be described here.

(2) Middle working area of left working chamber 1

As shown in fig. 10, the middle working area of the left working chamber 1 mainly comprises a turntable 15, a groove frame 16, a placing table 17, a consumable frame 19, a cell counter 111, a refrigerator 112, a dust detection module 113, a clamping jaw 114 and other devices which are arranged on a left working table plate 14, and can realize the procedures of storage, taking, uncovering, covering and cell counting of equipment such as cells and culture mediums filled in a culture flask 10, cell tubes, culture flask 10, culture medium bottles 20, centrifuge tubes, tip tubes and the like. The revolving stage 15 is established in left working chamber 1 and is leaned on the front position, and dust detection module 113, cell frame 16, place platform 17 clamping jaw 114 and fridge 112 are established in left working chamber 1's intermediate position, and consumable frame 19 and cell counter 111 are established in left working chamber 1's back position, and the overall arrangement is reasonable, and the automatic control and the mutually supporting of each device of being convenient for are mutually supported, and the operating efficiency is high.

Eight bottle clamp assemblies for placing the culture bottles 10 are mounted on the rotary base of the turntable 15, and a bottle cap seat 152 is arranged on one side of each bottle clamp assembly for storing the caps of the culture bottles 10. The rotation mechanism and principle of the turntable 15 are basically identical to those disclosed in the previous generation device CN115895894a, and are not described in detail herein, the main difference being that the structure of the bottle clamp assembly is optimized and improved.

As shown in fig. 10 and 11, the bottle clamp assembly mainly comprises an openable clamp 151, an electric clamp groove 153 for controlling the clamp 151, a support 154 and a steering engine 155, wherein the lower end of the steering engine 155 is fixedly arranged on a rotating base of the turntable 15, the support 154 is rotatably arranged at the upper end of the steering engine 155, the electric clamp groove 153 is arranged on the support 154, the clamp 151 is correspondingly arranged at the output end of the electric clamp groove 153, the electric clamp groove 153 is used for controlling the clamp 151 to open and close, and the steering engine 155 drives the clamp 151 and the electric clamp groove 153 to be unfolded and lifted. The back of the jig 151 is provided with a heating sheet 156.

The turntable 15 is mainly used for temporarily storing the culture flask 10 in a preliminary treatment process of cell culture and a cell harvesting process. The inner manipulator 25 installed on the right working chamber 2 clamps the culture flask 10 after sterilization (sterilization is completed in the sterilization mechanism 22 of the right working chamber 2), the corresponding rotary table 15 is matched with rotation to enable the flask clamping components to be set to the working positions one by one, the inner manipulator 25 places the culture flask 10 on the clamp 151, the clamp 151 is controlled by the electric clamp groove 153 to clamp the culture flask 10, the three-axis manipulator 18 of the upper working area of the left working chamber 1 uncaps the culture flask 10, then the filling and capping of cells and culture medium in the culture flask 10 are carried out, the culture flask 10 is horizontally placed, the culture flask 10 is moved out of the rotary table 15 to the culture box 4, and the pretreatment procedure of cell culture is completed. In the cell harvesting step, the culture flask 10 is moved out of the incubator 4 into the working chamber by the external manipulator 5, and the internal manipulator 25 cooperates with the turntable 15 to store, open and shake the culture flask 10, and then the triaxial manipulator 18 performs steps such as cell cleaning and pancreatin digestion.

As shown in FIG. 10, the sample loading assembly is formed by integrally mounting the trough frame 16, the placing table 17 and the clamping jaw 114 on a plate for temporarily storing the used tubes or bottles, optimizing the space arrangement and the operation logic, and completing the quantitative feeding of pancreatin, preservation solution, culture medium and the like into the culture flask 10 by matching with the internal manipulator 25.

In this embodiment the tank rack 16 is provided with three tank bottoms, the bottom of which is provided with heating plates. One of which is used for storing the culture medium bottle 20, and the other two of which can be used for storing physiological saline bottles or bottle bodies required by cell culture, and when certain cells are cultured, the culture medium needs to be kept at a constant temperature, and then the heating plate of the bottle groove can be started. The placing table 17 is used for placing the caps of various bottle bodies.

With reference to fig. 6, three clamping jaws 114 are provided in this embodiment, which can clamp both large-sized square straight-mouth bottles and small-sized round centrifuge tubes, so as to meet the requirements of various bottle bodies, or a person skilled in the art can select different numbers of clamping jaws or clamping jaws with other shapes and specifications.

As shown in fig. 6 and 10, the consumable rack 19 is of a rack structure, and is detachably mounted on the left working table 14, so that when consumable materials such as tip tubes and counting plates are required to be replenished, the switch door behind the left working chamber 1 can be opened, and the consumable rack 19 can be directly taken out, so that the consumable rack is quite convenient.

The cell counter 111 of this embodiment is a commercially available product, and is disposed beside the consumable rack 19, the suction cup of the three-axis mechanical arm 18 places the cell counting plate on the consumable rack 19 into the cell counter 111, so that cell counting can be quickly and conveniently implemented, and the structure and the usage mode of the cell counter 111 are not described herein in detail by combining the optical imaging technology and the intelligent image recognition technology and quantitatively analyzing the cell information in the acquired image by analyzing software.

(3) Upper working area of left working chamber 1

The upper working area of the left working chamber 1 mainly comprises a three-axis mechanical arm 18, a monitoring camera, a hepa system 6 and the like, wherein the monitoring camera and the hepa system 6 are all commercial products, so that the installation and the use of the monitoring camera and the hepa system are not repeated, an X-axis component and a Y-axis component of the three-axis mechanical arm 18 are basically universal structures in the prior art, and as shown in fig. 12, an execution end of the Z-axis component mainly comprises four parts, namely a sucker I181, a cover opening clamping jaw 182, a large plug 183 for taking a 10-m-p head and a small plug 184 for taking a 1-m-p head. The sucker I181 can be used for sucking a cell tube, a centrifuge tube or a cell counting plate and the like, and the action of the sucker I181 is flexibly set according to the cell culture process.

The left working table 14 is also provided with a waste bottle throwing port for discarding consumable materials such as centrifuge tubes, various bottle bodies, tip tubes and the like, and a waste bottle box is correspondingly arranged on the working table 14 or the lower working area of the left working chamber 1 in a matched manner. The waste bottle port may be located in a space adjacent to the sample addition assembly and consumable rack 19.

(II) Right working chamber 2

As shown in fig. 13 to 15, the right working chamber 2 has a similar external structure to the left working chamber 1: the whole is of a frame structure, the right outer cover 21 is externally installed, and the front surface, the rear surface and the right surface of the right outer cover 21 are respectively provided with switch doors for observation, debugging, overhaul and the like.

(1) Lower working area of right working chamber 2

The lower working area of the right working chamber 2 is mainly provided with a centrifuge 23, a distribution box and the like, and a plurality of storage areas for storing consumables such as vials and the like can be also arranged.

(2) Middle working area of right working chamber 2

As shown in fig. 13 to 15, the middle working area of the right working chamber 2 mainly includes a sterilizing mechanism 22 mounted on a right working table 24, an inner manipulator 25, a weighing scale 26, a vial rack 27, a waste liquid tank 28, a cover plate i 212, and the like. The devices in the middle working area of the right working chamber 2 are reasonably distributed, are convenient for automatic control and mutual matching, and have high working efficiency.

The sterilizing mechanism 22 and the cover plate I212 are positioned at the front position of the right working table plate 24, the sterilizing mechanism 22 is used for sterilizing the culture bottles 10, and the code scanner 213 is arranged on the sterilizing mechanism 22 and is used for scanning code records of the culture bottles 10 and the like; the cover plate I212 is used for being matched with the centrifugal machine 23, and the cover plate I212 can be opened and closed electrically.

The inner manipulator 25 and the waste liquid barrel 28 are arranged at the middle position of the right working table plate 24, the inner manipulator 25 can take into account the taking and placing of the culture bottle 10, the culture medium bottle 20, the centrifuge tube and other devices in the left working chamber 1 and the right working chamber 2, and the waste liquid barrel 28 is used for containing the waste culture liquid and other waste liquid in the culture bottle 10 after the cell culture is completed.

The weighing device 26 and the vial rack 27 are arranged at the rear position of the right working table plate 24, the weighing device 26 is used for weighing the weight of the culture medium vial 20 so as to avoid empty vial operation, the vial rack 27 is used for storing consumable materials such as the small centrifuge tube 30 and the large centrifuge tube 40, and when the consumable materials need to be supplemented, the opening and closing door at the rear of the right working chamber 2 can be opened to directly operate very conveniently.

As shown in FIGS. 16 and 17, the culture flask 10 may be placed in a sterilization mechanism 22 for sterilization, which includes an isolation bin 221, a cover plate II 222, a carrier 223 for placing the culture flask 10, a fan 224, and a spray head 225. The front side of the isolation bin 221 is provided with an opening for taking and placing the culture flask 10 by the outer manipulator 5, and the front side of the right outer cover 21 is correspondingly provided with an electric control bin door 211, so that the isolation bin 221 is ensured to be in a closed state during disinfection. The top of the isolation bin 221 is provided with an electrically-controlled opening and closing cover plate II 222, when the culture flask 10 is sterilized in the isolation bin 221, the cover plate II 222 is opened, and the inner manipulator 25 can take out the culture flask 10. The carrier 223 is installed in the isolation bin 221, the fan 224 and the two spray nozzles 225 are arranged at the rear side of the inside of the isolation bin 221 and are opposite to the carrier 223, the two spray nozzles 225 spray disinfectant (such as alcohol), and the fan 224 further sprays disinfectant to increase the disinfection area and the spraying uniformity.

(3) Upper working area of right working chamber 2

The upper working area of the right working chamber 2 is mainly equipped with a monitoring camera, another set of hepa systems 6, etc.

In the working chamber, the three-axis mechanical arm 18 is responsible for opening and closing cover actions, liquid sucking actions, sucking and grabbing cell counting plates, various bottle bodies and the like of all bottle tube equipment, and the inner mechanical arm 25 is responsible for clamping all bottle tube equipment. The external manipulator 5 is responsible for taking and placing the culture bottles 10 and the culture bottle trays.

As shown in fig. 18 and 19, the execution end of the outer manipulator 5 includes a tray clamp 51, a suction cup 52, a shield ii 53 and a mounting plate 57, the execution end is connected with the main body of the outer manipulator 5 through the mounting plate 57, a terminal clamp 54, a controller 55, an air pump 56 and the like are arranged in the shield ii 53, the terminal clamp 54 is driven by the main body of the outer manipulator 5 to realize the opening and closing clamp of the tray clamp 51, the air pump 56 is communicated with the suction cup 52 through a pipeline and a valve, the terminal clamp 54 and the controller 55 are respectively arranged at two sides of the mounting plate 57, and the air pump 56 and the pipeline, the valve and the like thereof are arranged on the inner side surface of the terminal clamp 54. The external manipulator can both grip the tray and hold the flask 10. The tray is used for arranging and storing the culture bottles 10, and the executing end of the outer manipulator 5 clamps the tray to conveniently enter and exit the incubator 4. When the operations such as coding, sterilizing and microscopic observation are needed for the culture flask 10, the sucker 52 sucks the culture flask 10 to execute corresponding actions, so that the process is concise, the efficiency is high, and the automatic control is convenient.

The device can be used for culturing, harvesting, analyzing and preserving human source samples such as adherent cells (mesenchymal cells, fibroblasts and tumor cells) and suspension cells (NK cells, CIK cells and Car-T cells). Taking a certain adherence type cell as an example, the automatic cell culture working process of the device comprises the following steps:

s1) pretreatment

1.1 coding: the conveying mechanism 121 stretches out, the outer mechanical arm 5 takes out the culture bottle 10 from the empty bottle frame 3 and puts the culture bottle 10 on the conveying mechanism 121, the conveying mechanism 121 conveys the culture bottle 10 into the protecting cover I122, the laser marking machine starts marking, and the conveying mechanism 121 conveys the culture bottle 10 out after the marking is finished;

1.2 disinfection: the outer mechanical arm 5 grabs the coded culture bottle 10 and puts the culture bottle 10 into the sterilizing mechanism 22 for sterilization, and the rear cover plate II 222 is opened;

in this embodiment, eight culture flasks 10 are used for each cell culture, and each culture flask 10 is subjected to coding and sterilization in sequence;

1.3 cover opening of flask 10: the inner manipulator 25 grabs the sterilized culture bottles 10 and sequentially places the culture bottles 10 on the bottle clamp assembly of the turntable 15, the three-axis manipulator 18 sequentially uncaps the culture bottles 10, and the cover is placed on the bottle cap seat 152;

1.4 pouring the culture solution: the internal manipulator 25 takes out the culture medium bottle 20, firstly, the solution in the culture medium bottle 20 is ensured to be enough by weighing in the weighing device 26, then the culture medium bottle 20 is placed on the groove frame 16, the three-axis manipulator 18 uncaps the culture medium bottle 20, and the internal manipulator 25 sequentially grabs the culture medium bottle 20 and pours the culture medium into the culture bottle 10;

1.5 cell separation flask: the three-axis mechanical arm 18 sucks the cell tube from the refrigerator 112 to be temporarily stored at the clamping jaw 114 and then uncaps, and the three-axis mechanical arm 18 sequentially sucks a certain amount of cells to be added into the culture flask 10;

1.6 flask culture: the three-axis mechanical arm 18 closes (closes) the culture flask 10, the inner mechanical arm 25 sequentially removes the culture flask 10, and the outer mechanical arm 5 sequentially places the culture flask 10 on the tray.

S2) cell culture

The external manipulator 5 places the tray into the incubator 4 to perform cell culture.

In the process of cell culture, the external manipulator 5 cooperates with the microscope assembly 13 to perform fusion degree analysis:

2.1, the incubator 4 is opened, the outer manipulator 5 takes out the tray and puts the tray on the empty bottle rack 3, and the incubator 4 is closed;

2.2 the conveyor carrier of the microscope assembly 13 extends out, the outer manipulator 5 places the culture flask 10 on the conveyor carrier, fusion analysis is carried out to the microscope, and after analysis, the outer manipulator 5 places the culture flask back to the tray.

Two or more culture bottles 10 are randomly extracted, if the fusion degree is not lower than 90%, the culture is qualified, cell harvest can be carried out, and if the fusion degree is lower than 90%, the outer manipulator 5 clamps the tray again and sends the tray back to the incubator 4 for continuous culture.

S3) cell harvesting

3.1, the incubator 4 is opened, the outer manipulator 5 takes out the tray and puts the tray on the empty bottle frame 3, and the incubator 4 is closed;

3.2 correction position: the conveying mechanism 121 stretches out, the external manipulator 5 takes out the culture bottle 10 from the tray and puts the culture bottle 10 on the conveying mechanism 121, and the external manipulator 5 corrects the culture bottle 10 to a required position;

3.3, disinfection: the outer mechanical arm 5 grabs the culture bottle 10 and puts the culture bottle 10 into the sterilizing mechanism 22 for sterilization, and the rear cover plate II 222 is opened;

3.3 placing flask 10: the inner manipulator 25 grabs the sterilized culture bottles 10, sequentially sweeps and stacks the culture bottles on the bottle clamp assembly of the turntable 15;

3.4 harvesting: the inner manipulator 25 takes out the small centrifuge tube 30 and the large centrifuge tube 40 and respectively puts them on the clamping jaw 114, the three-axis mechanical arm 18 sequentially uncaps the culture bottle 10, transfers the culture medium in the culture bottle 10 into the large centrifuge tube 40 with 250ml, and then carries out cell cleaning, pancreatin digestion (shaking), neutralization, cell collection, centrifugation, bottle closing, resuspension counting and the like, thus completing cell harvesting.

In the embodiment, the single batch culture time of the adherent cells by using the equipment and the process is 3-4 days, and the efficiency is obviously improved. Meanwhile, the cell culture process is only a preferred embodiment, and the device can complete automatic and efficient culture of various cells by setting corresponding program control components or action steps of mechanisms for different cell samples, different culture requirements and the like.

Claims (11)

1. An automated cell culture studio, characterized by: comprises a lower working area, a middle working area and an upper working area;

the lower working area is provided with a coding component (12), a microscope component (13), a centrifugal machine (23) and an electric component,

the middle working area is provided with a sterilizing mechanism (22), a turntable (15) for storing culture bottles (10), a vial sample adding assembly, a consumable rack (19), a cell counter (111), a refrigerator (112) for storing cells, an inner manipulator (25) and a vial rack (27);

the upper working area is provided with a triaxial mechanical arm (18) and a hepa system (6), and the triaxial mechanical arm (18) is used for being matched with the middle working area.

2. The automated cell culture working chamber of claim 1, wherein: the coding assembly (12), the microscope assembly (13) and the centrifugal machine (23) are sequentially arranged in front of the lower working area from left to right;

the inlet of the centrifugal machine (23) is communicated with the middle working area, and the inlets of the rotary table (15), the sterilizing mechanism (22) and the centrifugal machine (23) are sequentially arranged in front of the middle working area from left to right; the refrigerator (112), the vial loading assembly and the internal manipulator (25) are sequentially arranged in the middle position of the middle working area from left to right; the cell counter (111), the consumable rack (19) and the vial rack (27) are sequentially arranged at the rear position of the middle working area from left to right;

the triaxial mechanical arm (18) is matched with the left side area of the middle working area.

3. The automated cell culture working chamber of claim 1, wherein: the working chamber is divided into a left working chamber (1) and a right working chamber (2);

the left working chamber (1) is internally provided with a horizontal left working table plate (14) to form a separation of a lower working area and a middle working area, the coding assembly (12) and the microscope assembly (13) are positioned in front of the lower working area below the left working table plate (14), the rotary table (15) is positioned in front of the left working table plate (14), the refrigerator (112) and the vial sample adding assembly are positioned in the middle of the left working table plate (14), the cell counter (111) and the consumable rack (19) are positioned in the rear of the left working table plate (14), and the three-axis mechanical arm (18) is positioned in the upper working area of the left working chamber (1);

the utility model discloses a disinfection machine, including right working chamber (2), inner manipulator (25), pipe bottle frame (27) and pipe bottle frame (27), be equipped with horizontal right working platen (24) in right working chamber (2) and form the separation of lower part workspace and middle part workspace, centrifuge (23) are located the lower part workspace front right side of right working platen (24) below, the import of centrifuge (23) communicates with right working platen (24), the import of disinfection mechanism (22) and centrifuge (23) is located the front position of right working platen (24), interior manipulator (25) is located the median position of right working platen (24), pipe bottle frame (27) is located the rear position of right working platen (24).

4. The automated cell culture working chamber of claim 1, wherein: the marking assembly (12) comprises a laser marking machine, a protective cover I (122) and a conveying mechanism (121), wherein the laser marking machine comprises a machine base (123) and a marking head (124), the machine base (123) is positioned in the protective cover I (122), the conveying mechanism (121) is arranged on the machine base (123), an opening for passing laser of the marking head (124) is formed in the top of the protective cover I (122), and an opening for the telescopic in-out of the conveying mechanism (121) is formed in the front side of the protective cover I (122);

the top of the shield I (122) is provided with a reverse smoke pipe (125) and a fan (126), and the fan (126) is aligned to the reverse smoke pipe (125).

5. The automated cell culture working chamber of claim 1, wherein: the bottle clamp assembly comprises an openable clamp, an electric clamp groove for controlling the clamp, a support and a steering engine, wherein the lower end of the steering engine is fixedly arranged on a rotating base of the turntable, the support is rotatably arranged at the upper end of the steering engine, the electric clamp groove is arranged on the support, and the clamp is correspondingly arranged at the output end of the electric clamp groove.

6. The automated cell culture working chamber of claim 5, wherein: and a heating plate is arranged on the clamp.

7. The automated cell culture working chamber of claim 1, wherein: the vial loading assembly comprises a rack (16) for storing culture medium vials (20), a placement table (17) for storing caps, and a plurality of jaws for gripping the vials.

8. The automated cell culture working chamber of claim 1, wherein: the disinfection mechanism (22) comprises an isolation bin, an electrically-controlled opening and closing cover plate II, a carrier for placing the culture bottles (10), a fan and spray heads, wherein the front side of the isolation bin is provided with an opening, the cover plate II is positioned at the top of the isolation bin, the carrier is arranged in the isolation bin, and the fan and the two spray heads are arranged at the rear side of the inside of the isolation bin and are opposite to the carrier.

9. The automated cell culture working chamber of claim 1, wherein: the rear position of the middle working area is also provided with a weighing device (26) for weighing the culture medium bottles (20), the middle position of the middle working area is also provided with a waste liquid barrel (28), and the waste liquid barrel (28) is positioned on the right side of the inner manipulator (25).

10. An automatic change cell culture equipment, includes studio, empty bottle frame (3) several incubator (4) and outer manipulator (5), its characterized in that: the working chamber is the working chamber according to any one of claims 1 to 9.

11. The automated cell culture apparatus of claim 10, wherein: the outer manipulator (5) execution end comprises a tray clamp (51), a sucker (52), a shield II (53) and a mounting plate (57), the execution end is connected with the outer manipulator (5) main body through the mounting plate (57), and a tail end clamp holder (54), a controller (55) and an air pump (56) are arranged in the shield II (53).