Automated cell freezing and thawing equipment

Abstract

To provide a cell automatic freezing and thawing facility that can easily insert and remove cryotubes by a robot arm of an automatic transportation facility.SOLUTION: The cell automatic freezing and thawing facility includes an entrance / exit portion 1, a robot arm 2, a program freezer 3, a rack carrier 4, a cryopreservation tank 5, a carrier carrying-in / out mechanism 6, a plate transfer mechanism 7, a dry bath 8, a cell culture vessel temporary storage shelf 9, and a lid automatic opening / closing device 10. The rack carrier 4 includes a frame body and a plurality of cassette plates. The frame body includes a plurality of partition walls. A plurality of plate storage sections are provided between any two of the partition walls. Each of the plurality of cassette plates is separably connected to the plurality of plate storage sections while having a plurality of holding holes for holding cryotubes on each of the cassette plates.SELECTED DRAWING: Figure 10

Description

[Technical Field]

[0001] The present invention relates to biochemical laboratory equipment, and more particularly to an automatic cell freezing and thawing equipment. [Background technology]

[0002] In the cell preparation process, after undergoing the required processing, mass-cultured cells are dispensed into cryotubes and frozen in a cryopreservation tank (liquid nitrogen tank or gaseous nitrogen tank). To explain the cryopreservation method in detail, the cryotubes filled with cells are first placed in a programmable freezer to gradually cool the cells to -80°C. The cryotubes cooled to -80°C are then transferred to a tube shelf, and the entire tube shelf is then placed in a cryopreservation tank for cryopreservation. To thaw the cells, the entire tube shelf is first removed from the cryopreservation tank, the cryotubes filled with the cells to be thawed are removed from the tube shelf, and transferred to a thawing device where they are gently warmed and thawed. The thawed cells are then transferred from the cryotubes to a culture vessel filled with culture medium, completing the cell thawing process.

[0003] However, in the cell preparation process of the prior art, when cells are cryopreserved or thawed, the entire tube mounting shelf must be removed from the cryopreservation tank, forcing all of the cryopreserved cells to remain in a room temperature environment for a while. Furthermore, because the tube mounting shelf of the prior art is an integrally molded multi-layered frame-type shelf, it is difficult to insert and remove the cryotubes stored in the middle layer and inside the shelf using a robot arm of automated equipment. As a result, the operation of inserting and removing cryotubes from the tube mounting shelf of the prior art must be done manually. Therefore, the operation of inserting and removing cryotubes takes time and effort, and cells other than those to be removed are left in a room temperature environment for even longer, which may lead to unnecessary heating and adversely affect the cryopreservation of the cells. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] U.S. Patent Application Publication No. US2020 / 0114344 Summary of the Invention [Problem to be solved by the invention]

[0005] The present invention has been made in consideration of the drawbacks of the prior art, and aims to provide an automatic cell freezing and thawing equipment and a rack carrier to be used therewith, which has a frame body and cassette plate that are detachably connected, so that when inserting or removing a cryotube, the required cassette plate is removed from the frame body and the cryotube is inserted or removed.With this design, the robot arm of the automatic transport equipment can easily insert or remove the cryotube, making it possible to automate the freezing and thawing of cells. [Means for solving the problem]

[0006] In order to achieve the above object, the present invention proposes a rack carrier for use in an automatic cell freezing and thawing equipment, the rack carrier including a frame body and a plurality of cassette plates, the frame body includes a plurality of partition walls, and a plurality of plate accommodating portions are provided between any two of the partition walls; The plurality of cassette plates are detachably coupled to the plurality of plate accommodating portions, and each cassette plate has a plurality of holding holes for holding cryotubes.

[0007] a plurality of support ribs are provided on each of the partition walls, and the plurality of plate accommodating portions are formed by the opposing support ribs on the adjacent partition walls; Each of the cassette plates is detachably supported on a support rib of either of the two partition walls.

[0008] Each of the support ribs has a projection for preventing slip-out, and each of the cassette plates has two recesses for preventing slip-out, which receive the projections of the two support ribs.

[0009] The frame body has a central axis, the plurality of partition walls are spaced apart to surround the central axis and extend radially from the central axis, and each cassette plate is fan-shaped.

[0010] In order to achieve the above object, the present invention further proposes an automatic cell freezing and thawing equipment, which includes an access section, a robot arm, a programmable freezer, a rack carrier, a cryopreservation tank, a carrier loading / unloading mechanism, a plate transport mechanism, a dry bath, a cell culture vessel temporary storage shelf, and an automatic lid opening / closing device, the access part is used for the access of a cryotube or a cell culture vessel, the robot arm is used to transport the cryotube or the cell culture vessel; the programmable freezer is used to cool a cryotube carried in from the access section by the robot arm; The rack carrier includes a frame body and a plurality of cassette plates; the frame body includes a plurality of partition walls, and a plurality of plate accommodating portions are provided between any two of the partition walls; the plurality of cassette plates are detachably coupled to the plurality of plate accommodating portions, and each cassette plate has a plurality of holding holes for holding cryotubes; The cryopreservation tank has a cryopreservation space therein, the carrier carrying-in / out mechanism is used to carry the rack carrier into and out of the cryopreservation space of the cryopreservation tank; the plate transfer mechanism is used to place or remove one of the plurality of cassette plates into or from one of the plurality of plate accommodating units on the rack carrier that has been transferred from the cryopreservation space by the carrier transfer mechanism; the dry bath is used to heat the cryotube carried in from a cassette plate on the plate transfer mechanism by the robot arm; the cell culture vessel temporary storage shelf is used for temporarily storing the cell culture vessel carried in from the entrance / exit section by the robot arm, The automatic lid opening / closing device is characterized in that it is used to open and close the lids of cryotubes transported from the dry bath by the robot arm, and to open and close the lids of cell culture vessels transported from the temporary cell culture vessel storage shelf by the robot arm.

[0011] the automatic cell freezing and thawing equipment includes a refrigerated chamber whose internal temperature is lower than room temperature, and the rack carrier, the cryopreservation tank, and the carrier loading / unloading mechanism are arranged in the refrigerated chamber; The plate transfer mechanism is disposed outside the refrigerator compartment so as to be able to enter the refrigerator compartment, and is used to enter the refrigerator compartment and place or remove one of the plurality of cassette plates in one of the plurality of plate accommodating sections.

[0012] The automatic cell freezing and thawing equipment includes a low-temperature chamber that communicates with the refrigerating chamber and has a robot arm entrance opening on its wall, and an insulating lid that selectively opens and closes the robot arm entrance opening, the plate transport mechanism is disposed within the cryogenic chamber; The robot arm is characterized in that it can reach the cryotube of the cassette plate on the plate transfer mechanism through the robot arm entrance opening opened by the heat insulating lid. [Effects of the Invention]

[0013] The advantage of the present invention is that the frame body and cassette plate of the rack carrier are configured to be separable, and when in use, the rack carrier is transported out of the cryopreservation tank via the carrier transport mechanism, and then the required cassette plate is removed from the frame body via the plate transfer mechanism.This makes it possible to use a robotic arm to insert and remove cryotubes.In this way, the robotic arm can quickly and easily insert and remove cryotubes placed on the middle layer and inside the shelf.Furthermore, since the rack carrier can be returned into the cryopreservation tank first during the insertion and removal operation, the time it takes for cells other than those being removed to be removed from the cryopreservation tank is shortened, thereby effectively suppressing unnecessary temperature increases and preventing adverse effects on the cryopreservation of cells. [Brief explanation of the drawings]

[0014] [Figure 1] FIG. 1 is a perspective view of the automated cell freezing and thawing equipment of the present invention. [Figure 2] 1 is a perspective view of the automatic cell freezing and thawing equipment of the present invention, seen from the side where the refrigeration chamber is located. [Figure 3] FIG. 1 is a perspective view of the automated cell freezing and thawing equipment of the present invention, seen from the side where the low-temperature chamber is located. [Figure 4A] FIG. 2 is a top cross-sectional view showing the state in which a tube temporary storage rack is placed at the entrance / exit section of the automatic cell freezing / thawing equipment of the present invention. [Figure 4B] FIG. 2 is a top cross-sectional view showing a state in which a cell culture vessel is placed on an entrance / exit section of the automatic cell freezing / thawing equipment of the present invention. [Figure 5] 1 is a diagram showing the positional relationship between a rack carrier and a shelf plate transport mechanism according to the present invention; [Figure 6] FIG. 2 is an exploded perspective view of the cassette plate and the frame body of the present invention. [Figure 7] FIG. 2 is a perspective view showing a plate transfer mechanism of the present invention. [Figure 8] FIG. 1 is a perspective view showing a robot arm of the present invention. [Figure 9] FIG. 2 is an enlarged view of a local portion of the robot arm of the present invention. [Figure 10] FIG. 1 is a schematic diagram showing how cells are cryopreserved in the present invention. [Figure 11] FIG. 1 is a schematic diagram showing the process of thawing cells in accordance with the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0015] Hereinafter, the technical means adopted by the present invention to achieve the predetermined objects of the invention will be described in more detail with reference to the drawings and preferred embodiments of the present invention.

[0016] As shown in Figures 1, 4A, 4B and 10, the automatic cell freezing and thawing equipment of the present invention includes an access section 1, a robot arm 2, a programmable freezer 3, a rack carrier 4, a cryopreservation tank 5, a carrier loading / unloading mechanism 6, a plate transfer mechanism 7, a dry bath 8, a cell culture vessel temporary storage shelf 9, an automatic lid opening / closing device 10, a refrigerating chamber 11, a low-temperature chamber 12 and an insulated lid 13.

[0017] As shown in FIGS. 3, 4A, and 4B, the access section 1 is used for the entry and exit of a tube temporary storage rack B and a cell culture vessel C.

[0018] 10 and 11, during use, the cryotube A placed in the access section 1 is transferred to the programmable freezer 3 by the robot arm 2, and the temperature is gradually lowered in the programmable freezer 3. In addition, as shown in FIGS. 8 and 9, in a preferred embodiment of the present invention, the robot arm 2 preferably has a first gripper 21 that grips the cryotube A placed in the access section 1, and a second gripper 22 that grips the cell culture vessel C.

[0019] 5 and 6, the rack carrier 4 includes a frame body 41 and a plurality of cassette plates 42. The frame body 41 has a plurality of partition walls 411, and a plurality of plate accommodating sections 412 are provided between any two of the partition walls 411, and the plurality of cassette plates 42 are respectively detachably coupled to the plate accommodating sections 412. Each cassette plate 42 has a plurality of tube holding holes 421 for accommodating cryotubes A, and the cryotubes A are transferred into the tube holding holes 421 by the robot arm 2 and accommodated therein.

[0020] In this embodiment, a plurality of support ribs 413 are provided on each partition wall 411, and the plurality of plate accommodating sections 412 are composed of a plurality of support ribs 413 on one of the partition walls 411 and a plurality of aligned support ribs 413 on the other adjacent partition wall 411, and each cassette plate 42 is supported on the support ribs 413 between any two adjacent partition walls 411. Also, each support rib 413 has a fall-out prevention protrusion 414, and each cassette plate 42 has two corresponding fall-out prevention recesses 422 that receive the fall-out prevention protrusions 414, and this configuration prevents the cassette plates 42 supported on the support ribs 413 from accidentally falling off. Furthermore, in this embodiment, the frame main body 41 has a central axis 415, and the plurality of partition walls 411 are arranged at intervals so as to surround the central axis 415 of the frame main body 41, and extend radially of the central axis 415. It is preferable that the cassette plate 42 is a fan-shaped plate, but this is not limited to this. In addition, the joining structure between the frame main body 41 and the cassette plate 42 is not limited to the structure described above, and it is sufficient that the cassette plate 42 and the frame main body 41 can be combined in a separable manner.

[0021] The cryopreservation tank 5 has a cryopreservation space inside.

[0022] The carrier transfer mechanism 6 is used to transfer the rack carrier 4 into and out of the cryopreservation tank 5 .

[0023] The plate transfer mechanism 7 is used to place the cassette plate 42 in one of the multiple plate storage units 412 on the rack carrier 4 that has been transported out of the cryopreservation space of the cryopreservation tank 5 by the carrier transport mechanism 6, or to remove the required cassette plate 42 from the transported plate storage unit 412. Specifically, as shown in FIGS. 5 and 7 , the plate transfer mechanism 7 includes a rail member 71 and a clamping member 72 that is movable on the rail member 71 and can clamp the cassette plate 42. When the plate transfer mechanism 7 moves a cassette plate 42 in or out, the clamping member 72 moves along the rail member 71 toward the rack carrier 4 that has been transported out of the cryopreservation space by the carrier transport mechanism 6, and places the cassette plate 42 in the plate storage unit 412 of the rack carrier 4, or removes the cassette plate 42 from the plate storage unit 412 of the rack carrier 4. Note that the specific structure of the plate transfer mechanism 7 is not limited to the above-described configuration.

[0024] The dry bath 8 is used to gradually increase the temperature of the cryotube A that has been transferred from the cassette plate 42 on the plate transfer mechanism 7 by the robot arm 2 .

[0025] The cell culture vessel temporary storage shelf 9 is used to temporarily store the cell culture vessel C transferred from the access section 1 by the robot arm 2.

[0026] The automatic lid opening / closing device 10 is used to open and close the lids of cryotubes A that have been transferred from the dry bath 8 by the robot arm 2, and the lids of cell culture vessels C that have been transferred from the cell culture vessel temporary storage shelf 9 by the robot arm 2.

[0027] 1 to 4A and 4B, an automated cell freezing and thawing equipment according to a preferred embodiment of the present invention includes a refrigeration compartment 11, a low-temperature chamber 12, and an insulated lid 13. The internal temperature of the refrigerated space of the refrigeration compartment 11 is preferably about 4 degrees Celsius lower than the external temperature. The low-temperature chamber 12 is installed to communicate with the refrigeration compartment 11 and functions as an extension of the refrigerated space of the refrigeration compartment 11. A robot arm access opening 121 is installed on the wall of the low-temperature chamber 12. The insulated lid 13 is installed at the installation position of the robot arm access opening 121 and selectively opens and closes the robot arm access opening 121.

[0028] In this embodiment, the rack carrier 4, cryopreservation tank 5, and carrier loading / unloading mechanism 6 are disposed within the refrigeration chamber 11. The plate transfer mechanism 7 is disposed within the low-temperature chamber 12 and is capable of entering the refrigeration chamber 11. More specifically, the plate transfer mechanism 7 enters the refrigeration chamber 11 through a communication passage between the low-temperature chamber 12 and the refrigeration chamber 11 to place or remove one of the cassette plates 42 in one of the plate receiving sections 412. The robot arm 2 passes through the robot arm entrance opening 121 opened by the insulating lid 13, moves to the cassette plate 42 placed on the plate transfer mechanism 7, and deposits a cryotube A in the tube holding hole 421 of the cassette plate 42, or removes one of the cryotubes A received in the tube holding hole 421.

[0029] By installing the refrigeration chamber 11, low-temperature chamber 12 and insulated lid 13, the rack carrier 4, the cassette plate 42 and the cryotube A placed on it are not directly exposed to the room temperature environment after being removed from the cryopreservation tank 5, but remain in a low-temperature space with a relatively low temperature, thereby improving the cryopreservation stability of cells.

[0030] A series of automatic loading and unloading operations of the automatic cell freezing and thawing equipment according to a preferred embodiment of the present invention will now be described.

[0031] First, the cell cryopreservation process will be described with reference to FIGS. 4A, 6 and 10. FIG.

[0032] After injecting a cell solution to be cryopreserved into a cryotube A, which has RFID (Radio Frequency Identification) identification information pre-written therein, the cryotube A is placed in a temporary tube storage rack B and placed in the access section 1. As shown in FIGS. 9 and 10 , the robot arm 2 then reads the RFID identification information using its RFID reader 23, removes the specific cryotube A from the temporary tube storage rack B, and transfers it to the programmable freezer 3 for a first cooling cycle. Meanwhile, the temporary tube storage rack B from which the cryotube A was removed is retrieved from the access section 1. After the first cooling cycle of the cryotube A is completed, the lid of the cryopreservation tank 5 is opened, the carrier loading / unloading mechanism 6 removes the rack carrier 4 from the cryopreservation tank 5, and the plate transfer mechanism 7 removes one of the multiple cassette plates 42 housed in the rack carrier 4 from the frame main body 41. The robot arm 2 then transfers the cryotube A, whose first cooling cycle has been completed, to the cassette plate 42 previously removed by the plate transfer mechanism 7. Finally, the plate transfer mechanism 7 returns the cassette plate 42 carrying the cryotube A to the plate storage section 412 of the frame main body 41, and the carrier transport mechanism 6 transports the rack carrier 4 into the cryopreservation space of the cryopreservation tank 5, and the lid of the cryopreservation tank 5 is closed, completing the cell cryopreservation process.

[0033] Furthermore, the cell thawing process will be explained with reference to FIG. 4B and FIG.

[0034] Before thawing the cells, RFID identification information is written in advance to the cell culture vessel C, and a cell culture solution is injected into the cell culture vessel C via an external dispensing device (the dispensing device is not included in the present invention). The cell culture vessel C is then placed in the access section 1 and transferred to a temporary cell culture vessel storage shelf 9 by a robot arm 2, where it is kept waiting. Referring to FIG. 11 , when thawing the cells, the robot arm 2 first removes the cell culture vessel C from the temporary cell culture vessel storage shelf 9 and transports it to an automatic lid opening / closing device 10, where it is kept waiting, in order to open the lid of the cell culture vessel C. At the same time, the lid of the cryopreservation tank 5 is opened, and the carrier loading / unloading mechanism 6 removes the rack carrier 4 from the cryopreservation tank 5 by lifting it up. The plate transfer mechanism 7 removes from the frame body 41 a cassette plate 42 containing cryotubes A filled with the cell solution to be thawed, which are on the rack carrier 4. The robot arm 2 then reads the RFID identification information and transfers the cryotubes A to a dry bath 8 for thawing. Once a specific cryotube A has been transferred to the dry bath 8, the plate transfer mechanism 7 returns the cassette plate 42 to the frame body 41, and the carrier loading / unloading mechanism 6 transports the rack carrier 4 into the cryopreservation tank 5, after which the lid of the cryopreservation tank 5 is closed. Next, after the cell solution in the cryotube A has thawed, the robot arm 2 transports the cryotube A to the automatic lid opener 10 to open the lid of the cryotube A. After the lids of both the cell culture vessel C and the cryotube A transported to the automatic lid opener 10 have been opened, the thawed cell solution in the cryotube A is injected into the cell culture vessel C, and the lid of the cell culture vessel C is closed again via the automatic lid opener 10. Finally, the robot arm 2 reads the RFID identification information of the cell culture vessel C into which the cell solution has been injected, and moves the cell culture vessel C to the entrance / exit section 1, completing the cell thawing operation, while the cryotube A into which the cell solution has been discharged is disposed of by being thrown into a waste collection box 14 after its lid is closed. The cell culture vessel C into which the cell solution has been injected and which has been transferred to the entrance / exit section 1 is transported to a cell culture mechanism (the transport means and the cell culture mechanism are not included in the present invention) via an external transport means, where the subsequent cell culture is carried out.

[0035] The advantage of the present invention is that the frame body 41 and cassette plate 42 of the rack carrier 4 are configured to be separable, and when in use, the rack carrier 4 is transported out of the cryopreservation tank 5 via the carrier transport mechanism 6, and then the required cassette plate 42 is removed from the frame body 41 via the plate transfer mechanism 7. This makes it possible to use the robot arm 2 to insert and remove cryotubes A. In this way, the robot arm 2 can quickly and easily insert and remove cryotubes A placed on the middle layer and inside the shelf. Furthermore, since the rack carrier 4 can be returned into the cryopreservation tank 5 first during the insertion and removal operation, the time it takes for cells other than those being removed to be removed from the cryopreservation tank 5 is shortened, thereby effectively suppressing unnecessary temperature rise and preventing adverse effects on the cryopreservation of cells.

[0036] The above description is merely a preferred embodiment of the present invention and does not limit the present invention in any way. Although the present invention has been disclosed above with a relatively preferred embodiment, this does not limit the present invention, and any simple modifications, changes and alterations made by anyone skilled in the art to the above embodiment based on the essence of the technology of the present invention, within the scope of the technical concept of the present invention, still fall within the scope of the technical concept of the present invention. [Explanation of symbols]

[0037] A Cryotube B Tube temporary storage rack C Cell culture container 1 Entrance and Exit 2. Robotic Arm 21 First gripper 22 Second gripper 23 RFID reader 3 Program Freezer 4 Rack Carriers 41 Frame body 411 Partition Wall 412 Plate storage section 413 Support Rib 414 Anti-slip protrusion 415 Center axis 42 Cassette Plate 421 Tube holding hole 422 Recessed hole to prevent slipping out 5. Cryopreservation tank 6 Carrier loading / unloading mechanism 7 Plate transfer mechanism 71 Rail components 72 Clamping member 8 Dry Baths 9 Cell culture container temporary storage shelf 10 Automatic lid opening and closing device 11 Refrigerator 12 Low temperature chamber 121 Robot arm entrance opening 13 Insulated lid 14 Garbage collection bin

Claims

1. An automatic cell freezing and thawing facility, comprising: an access section, a robot arm, a programmable freezer, a rack carrier, a cryopreservation tank, a carrier loading / unloading mechanism, a plate transfer mechanism, a dry bath, a cell culture vessel temporary storage shelf, and an automatic lid opening / closing device; the access part is used for the access of a cryotube or a cell culture vessel, the robot arm is used to transport the cryotube or the cell culture vessel; the programmable freezer is used to cool a cryotube carried in from the access section by the robot arm; the rack carrier includes a frame body and a plurality of cassette plates, the frame body having a plurality of partition walls, a plurality of plate accommodating sections provided between any two of the partition walls, the plurality of cassette plates being detachably coupled to the plurality of plate accommodating sections, and each cassette plate having a plurality of holding holes for holding cryotubes; The cryopreservation tank has a cryopreservation space therein, the carrier carrying-in / out mechanism is used to carry the rack carrier into and out of the cryopreservation space of the cryopreservation tank; the plate transfer mechanism is used to place or remove one of the plurality of cassette plates into or from one of the plurality of plate accommodating units on the rack carrier that has been transferred from the cryopreservation space by the carrier transfer mechanism; the dry bath is used to heat the cryotube carried in from a cassette plate on the plate transfer mechanism by the robot arm; the cell culture vessel temporary storage shelf is used for temporarily storing the cell culture vessel carried in from the entrance / exit section by the robot arm, The automatic lid opening / closing device is used to open and close the lids of cryotubes transported from the dry bath by the robot arm, and to open and close the lids of cell culture vessels transported from the temporary cell culture vessel storage shelf by the robot arm.

2. the automatic cell freezing and thawing equipment includes a refrigerated chamber whose internal temperature is lower than room temperature, and the rack carrier, the cryopreservation tank, and the carrier loading / unloading mechanism are arranged in the refrigerated chamber; 2. The automatic cell freezing and thawing equipment according to claim 1, wherein the plate transfer mechanism is disposed outside the refrigeration chamber so as to be able to enter the interior of the refrigeration chamber, and is used to enter the interior of the refrigeration chamber and place or remove one of the plurality of cassette plates into one of the plurality of plate storage sections.

3. The automatic cell freezing and thawing equipment includes a low-temperature chamber that communicates with the refrigerating chamber and has a robot arm entrance opening on a wall thereof, and an insulating lid that selectively opens and closes the robot arm entrance opening; the plate transport mechanism is disposed within the cryogenic chamber; 3. The automatic cell freezing and thawing equipment according to claim 2, wherein the robot arm can reach the cryotube of the cassette plate on the plate transfer mechanism through the robot arm access opening opened by the insulating lid.

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