Single cell cloning and picking device

Abstract

The utility model relates to the related technical field of single cell picking devices, in particular to a single cell clone picking device, which is used for picking single cells in a cell culture dish, the cell picking culture dish is positioned in a picking area of an inverted microscope objective table, and the single cell clone picking device is used for picking the single cells in the cell culture dish. Microscope collecting lenses with fixed positions are arranged above the picking area at intervals, and microscope collecting lenses are arranged above the picking area of the inverted microscope objective table at intervals; cells at the bottom of the culture dish are scraped off through the cell suction tube opening, the cells are sucked and stored in the cell suction tube by adjusting the position of the rubber plug through the suction adjusting assembly, the sucked cells can be pushed into a new culture dish containing a culture medium, and picking and transferring of single cell clones are achieved. The device is easy to assemble and disassemble, and is suitable for most inverted microscopes, and the cell suction tube opening covers single cell cloning, so that other parenchyma cells cannot be doped in the picking process, and the unicity of single cell cloning is ensured.

Description

Technical Field

[0001] The utility model relates to the technical field related to single cell picking devices, and more specifically, to a single cell clone picking device. Background Art

[0002] Single-cell cloning technology plays an important role in modern biomedical research and applications. The core of single-cell cloning technology is to isolate and select single-cell growing and proliferating cell clusters from complex cell populations, and then expand them in vitro to obtain cell populations with the same genetic background and functional characteristics.

[0003] With the development of technology, automated and intelligent instruments and equipment have been gradually applied to the acquisition of single cells, such as flow cytometry, microfluidics, laser capture microdissection, etc., but the equipment required for these technologies is expensive and the cost is high, which limits their large-scale use. Conventional methods for obtaining single-cell clones, such as limiting dilution, embryo mouth pipette picking, and pipette picking, although low in cost, each has its own shortcomings. The limiting dilution method is time-consuming, inefficient, and it is not easy to obtain a pure single-cell population. Embryo mouth pipette picking and pipette picking require the operator to use a handheld device to absorb cells, so the operator's technical requirements are relatively high. Although it is easy to absorb impure cells, it is difficult to obtain pure single cells. Therefore, a low-cost, easy-to-operate device is extremely necessary for the acquisition of single-cell clones. Utility Model Content

[0004] The purpose of the utility model is to provide a single cell clone picking device which is simple to operate and can easily obtain pure single cells.

[0005] To achieve the above-mentioned object, the present invention provides the following technical solutions: a single-cell clone picking device, the picking device is used to pick single cells from a cell culture dish, the picked cell culture dish is located in a picking area of ​​an inverted microscope stage, and a fixed microscope condenser is spaced above the picking area;

[0006] The picking device includes a picking mechanism and a supporting mechanism;

[0007] The picking mechanism includes a cell pipette, an injection tube, a rubber stopper, and a suction adjustment component; the cell pipette is connected to the support mechanism, and one end of the cell pipette is provided with a cell pipette port, which is used to scrape cells from the bottom of the culture dish; the other end of the cell pipette is connected to the connecting port of the injection tube through a rubber hose, and a rubber stopper is provided in a sealing and sliding manner at the end of the injection tube away from the connecting port, the outer wall of the rubber stopper contacts the inner wall of the injection tube and seals the injection tube, and the rubber stopper is also connected to the suction adjustment component, which absorbs the scraped cells by adjusting the position of the rubber stopper in the injection tube;

[0008] The support mechanism includes a first connecting component and a second connecting component. The first connecting component is detachably fixedly connected to the microscope condenser, and the cell pipette is detachably installed on the first connecting component. The angle of the cell pipette on the first connecting component is adjustable; the second connecting component is detachably fixedly connected to the inverted microscope stage; and the injection tube is fixedly installed on the second connecting component.

[0009] A further technical solution of the present application is: the suction adjustment assembly includes a first screw and an adjustment knob, the first screw is threadedly connected to the threaded inner wall of the injection tube, the two ends of the first screw are respectively connected to the rubber stopper and the adjustment knob, and the adjustment knob is located outside the injection tube.

[0010] A further technical solution of the present application is that the cell pipette port is arranged vertically.

[0011] A further technical solution of the present application is: the first connecting assembly includes a first clamp, a second clamp, a lifting rod, a connecting seat and two connecting parts, the first clamp is provided with a first clamping area for clamping a microscope condenser and a second clamping area for clamping the connecting seat, the lifting rod is installed on the connecting seat, the second clamp is installed at the end of the lifting rod through one of the connecting parts, and the second clamp is used to clamp the cell pipette.

[0012] A further technical solution of the present application is as follows: the lifting rod is inserted into a slot of the connecting seat and is slidingly connected to the connecting seat; a toothed portion is provided on one side of the lifting rod, the toothed portion is meshed with a gear, and the gear is rotatably connected to the connecting seat.

[0013] A further technical solution of the present application is: the first clamp includes two metal sheets, one end of the two metal sheets is fixedly connected by a connecting piece, and the other end of the two metal sheets clamps the microscope condenser and is fixed to the connecting seat by the connecting piece.

[0014] A further technical solution of the present application is that the clamping surface of the metal sheet is provided with anti-slip and anti-collision rubber.

[0015] A further technical solution of the present application is: the connecting piece includes a connecting protrusion, a polished rod and a threaded sleeve, one end of the polished rod is connected to the connecting protrusion, and the other end of the polished rod is provided with a threaded portion, and the threaded portion is threadedly connected to the threaded sleeve.

[0016] A further technical solution of the present application is: the second clamp includes two metal sheets, and the third clamping area for clamping the cell pipette is between the two metal sheets. The two ends of the two metal sheets are fixed by connecting parts, and one of the optical rods passes through one end of the two metal sheets in sequence and is threadedly connected to one of the threaded sleeves; the other optical rod passes through the other end of the two metal sheets and the lifting rod in sequence and is threadedly connected to the other threaded sleeve.

[0017] A further technical solution of the present application is: the second connecting component includes a connecting seat and a fixing clamp, the two ends of the connecting seat are fixedly connected to the injection tube and the fixing clamp respectively, and the fixing clamp is clamped and set on the inverted microscope stage.

[0018] Compared with the prior art, the technical solution provided by this utility model has the following beneficial effects:

[0019] The utility model stably mounts the cell pipette on the condenser of the inverted microscope by arranging a first clamp, then uses the cell pipette mouth to scrape off cells on the bottom of the culture dish, and adjusts the position of the rubber stopper by the suction adjustment component to suck the cells and store them in the cell pipette, so that the sucked cells can be pushed into a new cell culture dish containing culture medium, thereby realizing the picking and transfer of single-cell clones; the device is easy to assemble and disassemble and is suitable for most inverted microscopes, and the cell pipette mouth covers the single-cell clone, ensuring that other miscellaneous cells will not be mixed in during the picking process; at the same time, the threaded suction adjustment ensures that the suction is relatively stable when adjusting the suction, and will not suddenly increase or decrease, ensuring that other surrounding cells will not be sucked in when sucking single-clone cells, thereby ensuring the uniqueness of the single-cell clone. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a schematic structural diagram of a single cell clone picking device provided by the present invention;

[0021] Figure 2 This is a schematic structural diagram of a connector in a single-cell clone picking device provided by the present invention;

[0022] Figure 3 This is a schematic diagram of the structure of a cell pipette port in a single-cell clone picking device provided by the present invention;

[0023] Figure 4 This is a cross-sectional view of an injection tube in a single-cell clone picking device provided by the present invention;

[0024] Figure 5 This is a schematic diagram of the structure of the connection between the lifting rod and the cell pipette in a single-cell clone picking device provided by the present invention;

[0025] Figure 6 This is a schematic structural diagram of the connection between a microscope condenser and a connector in a single-cell clone picking device provided by the present invention;

[0026] Figure 7 This is a structural schematic diagram of a lifting rod and gears in a single-cell clone picking device provided by the utility model.

[0027] Explanation of the numbers in the schematic diagram:

[0028] 1. Microscope condenser; 2. First clamp; 3. Connector; 4. Anti-drop part; 5. Lifting rod; 6. Second clamp; 7. Cell pipette; 8. Rubber hose; 9. Connecting port; 10. Injection tube; 11. Connecting seat; 12. Adjusting knob; 13. Fixing clamp; 14. Inverted microscope stage; 15. Cell pipette port; 16. Cell culture dish; 17. Rubber stopper; 18. Second screw; 19. First screw; 20. Threaded tube; 21. Gear; 22. Connecting seat; 31. Connecting protrusion; 32. Optical rod; 33. Threaded part; 34. Threaded sleeve. DETAILED DESCRIPTION

[0029] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. The following is a further description of the present invention in combination with the embodiments.

[0030] See also Figures 1 to 7 In one embodiment of the present application, a single cell clone picking device is provided, which is used to pick single cells from a cell culture dish 16. The picked cell culture dish 16 is located in a picking area of ​​an inverted microscope stage 14. A fixed microscope condenser 1 is spaced apart above the picking area.

[0031] The picking device includes a picking mechanism and a supporting mechanism;

[0032] The picking mechanism includes a cell pipette 7, an injection tube 10, a rubber stopper 17 and a suction adjustment component; the cell pipette 7 is connected to the support mechanism, and one end of the cell pipette 7 is provided with a cell pipette port 15, and the cell pipette port 15 is used to scrape cells from the bottom of the culture dish; the other end of the cell pipette 7 is connected to the connecting port 9 of the injection tube 10 through a rubber hose 8, and the end of the injection tube 10 away from the connecting port 9 is sealed and slidably provided with a rubber stopper 17, the outer wall of the rubber stopper 17 contacts the inner wall of the injection tube 10 and seals the injection tube 10, and the rubber stopper 17 is also connected to the suction adjustment component, and the suction adjustment component absorbs the scraped cells by adjusting the position of the rubber stopper 17 in the injection tube 10;

[0033] The support mechanism includes a first connecting component and a second connecting component. The first connecting component is detachably fixedly connected to the microscope condenser 1, and the cell pipette 7 is detachably installed on the first connecting component. The angle of the cell pipette 7 on the first connecting component is adjustable; the second connecting component is detachably fixedly connected to the inverted microscope stage 14; the injection tube 10 is fixedly installed on the second connecting component.

[0034] In this embodiment, the first connecting assembly includes a first clamp 2, a second clamp 6, a lifting rod 5, a connecting seat 22 and two connecting parts 3. The first clamp 2 is provided with a first clamping area for clamping the microscope condenser 1 and a second clamping area for clamping the connecting seat 22. The lifting rod 5 is installed on the connecting seat 22. The second clamp 6 is installed at the end of the lifting rod 5 through one of the connecting parts 3. The second clamp 6 is used to clamp the cell pipette 7.

[0035] In order to facilitate the adjustment of the height of the cell pipette 7, in this embodiment, the lifting rod 5 is inserted into the slot of the connecting seat 22 and is slidably connected to the connecting seat 22. A toothed portion is provided on one side of the lifting rod 5, which is engaged with the gear 21, and the gear 21 is rotatably connected to the connecting seat 22.

[0036] In order to prevent it from falling, an anti-falling portion 4 is provided on the top of the lifting rod 5 .

[0037] Specifically, the first clamp 2 includes two metal sheets, one end of the two metal sheets is fixedly connected by a connector 3 , and the other end of the two metal sheets clamps the microscope condenser 1 and is fixed to the connecting seat 22 through the connector 3 .

[0038] In order to play a protective role, the clamping surface of the metal sheet is provided with anti-slip and anti-collision rubber.

[0039] In one case of this embodiment, the connecting member 3 includes a connecting protrusion 31, a polished rod 32 and a threaded sleeve 34, one end of the polished rod 32 is connected to the connecting protrusion 31, and the other end of the polished rod 32 is provided with a threaded portion 33, and the threaded portion 33 is threadedly connected to the threaded sleeve 34.

[0040] During actual use, one end of the polished rod 32 passes through the two metal sheets and is then put on the threaded sleeve 34 , and the two metal sheets are fixed by tightening the threaded sleeve 34 .

[0041] The second clamp 6 includes two metal sheets, and the third clamping area for clamping the cell pipette 7 is between the two metal sheets. The two ends of the two metal sheets are fixed by a connecting piece 3. One of the polished rods 32 passes through one end of the two metal sheets in sequence and is threadedly connected to one of the threaded sleeves 34; the other polished rod 32 passes through the other end of the two metal sheets and the lifting rod 5 in sequence and is threadedly connected to the other threaded sleeve 34.

[0042] In this embodiment, the second connecting component includes a connecting seat 11 and a fixing clamp 13 . Both ends of the connecting seat 11 are fixedly connected to the injection tube 10 and the fixing clamp 13 , respectively. The fixing clamp 13 is clamped on an inverted microscope stage 14 .

[0043] In order to play a protective role, the clamping surface of the fixing clamp 13 is provided with anti-slip and anti-collision rubber.

[0044] This embodiment is implemented as follows: first, place the cell culture dish 16 on the inverted microscope stage 14, use a 4x objective lens, and adjust the focal length; at the same time, connect the cell pipette 7 to the connecting port 9 through the rubber hose 8, and then fix the injection tube 10 on the connecting seat 11, and then clamp the fixing clamp 13 on the inverted microscope stage 14 to fix the position of the injection tube 10, first screw on the threaded sleeve 34 at the first clamping area to fix the two metal sheets, place the microscope condenser 1 in the first clamping area, place one end of the connecting seat 22 in the second clamping area, and then screw on the threaded sleeve 34 at the second clamping area to fix the microscope condenser 1 and the connecting seat 22; after adjusting the extension length of the lifting rod 5 by rotating the gear 21, One of the polished rods 32 sequentially passes through one end of the two metal sheets and is then threadedly connected to one of the threaded sleeves 34 to fix the two metal sheets. Then, the cell pipette 7 is placed in the third clamping area. Another polished rod 32 sequentially passes through the other end of the two metal sheets and the lifting rod 5 and is then threadedly connected to another threaded sleeve 34 to achieve installation of the cell pipette 7 on the lifting rod 5. After unscrewing out the other threaded sleeve 34 located on the lifting rod 5, the angle of the cell pipette 7 can be adjusted. After adjusting the angle, the other threaded sleeve 34 is tightened to fix the angle of the cell pipette 7. After unscrewing out one of the threaded sleeves 34 located on the two metal sheets, the position of the cell pipette 7 can be adjusted in the lateral direction. After adjusting the position, one of the threaded sleeves 34 is tightened to fix the position of the cell pipette 7.

[0045] During the operation, the cell pipette 7 is adjusted and observed through the microscope condenser 1 to ensure that the cell pipette port 15 of the cell pipette 7 covers the cell clone and is close to the bottom of the cell culture dish 16 without pressing it to ensure that the cell culture dish 16 can move, indicating that the angle of the cell pipette 7 is properly adjusted. Then, by moving the inverted microscope stage 14 to drive the cell culture dish 16, or directly moving the cell culture dish 16 manually, the cell pipette port 15 is used to scrape the cells from the bottom of the cell culture dish 16. The adjustment knob 12 is then rotated to drive the rubber stopper 17 in a direction away from the connection port 9. Under the action of negative pressure, the scraped cells are sucked into the cell pipette 7. The lifting rod 5 is then raised to move the cell pipette 7 upward, and a new cell culture dish 16 containing culture medium is replaced. The cell pipette 7 is then lowered and the adjustment knob 12 is reversed to push the monoclonal cells into the new cell culture dish 16. If the cells are tightly attached to the wall, trypsin can be used for digestion before picking.

[0046] In this embodiment, the suction adjustment assembly includes a first screw 19 and an adjusting knob 12. The first screw 19 is threadedly connected to the threaded inner wall of the injection tube 10. The two ends of the first screw 19 are respectively connected to the rubber stopper 17 and the adjusting knob 12. The adjusting knob 12 is located outside the injection tube 10 to facilitate adjusting the position of the rubber stopper 17.

[0047] In order to increase the movement stroke of the adjusting rubber stopper 17, a threaded tube 20 is provided at one end of the rubber stopper 17 close to the first screw 19, and a second screw 18 is provided at one end of the first screw 19 close to the rubber stopper 17. The second screw 18 is inserted into the threaded tube 20 and threadedly connected to the threaded tube 20.

[0048] The double-threaded knob design makes operation simple and the suction force stable, ensuring that other complex cells are not inhaled during the cell aspiration process, ensuring the purity of single-cell cloning.

[0049] To facilitate single cell extraction, in this embodiment, the cell pipette port 15 is vertically positioned. The cell pipette port 15 is bent and then vertically positioned to ensure that the cell clone is completely covered and isolated from other cells without affecting the observation field under the microscope condenser 1, thereby facilitating the extraction of cell clones.

[0050] In this embodiment, the cell pipette port 15 covers the single cell clone, ensuring that other foreign cells will not be mixed in during the picking process. At the same time, the threaded suction adjustment makes the suction relatively stable when adjusting the suction, and will not suddenly increase or decrease, ensuring that when a single clone cell is aspirated, other surrounding cells will not be aspirated, thereby ensuring the uniqueness of the single cell clone.

[0051] The above is a schematic description of the present invention and its embodiments, which is not restrictive. The drawings show only one embodiment of the present invention, and the actual structure is not limited thereto. Therefore, if a person skilled in the art is inspired by this and, without departing from the inventive purpose of the present invention, designs a structure and embodiment similar to the technical solution without inventiveness, they shall fall within the scope of protection of the present invention.

[0052] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art.

Claims

1. A single cell clone picking device, the picking device is used to pick single cells in a cell culture dish (16), the picked cell culture dish (16) is located in a picking area of ​​an inverted microscope stage (14), and a fixed microscope condenser (1) is spaced above the picking area; characterized in that The picking device includes a picking mechanism and a supporting mechanism; The picking mechanism comprises a cell pipette (7), an injection tube (10), a rubber stopper (17) and a suction adjustment component; the cell pipette (7) is connected to the supporting mechanism, one end of the cell pipette (7) is provided with a cell pipette port (15), and the cell pipette port (15) is used to scrape off cells on the bottom of the culture dish; the other end of the cell pipette (7) is communicated with the connecting port (9) of the injection tube (10) through a rubber hose (8), and a rubber stopper (17) is provided in a sealing and sliding manner in the end of the injection tube (10) away from the connecting port (9), the outer wall of the rubber stopper (17) contacts the inner wall of the injection tube (10) and seals the injection tube (10), and the rubber stopper (17) is also connected to the suction adjustment component, and the suction adjustment component absorbs the scraped cells by adjusting the position of the rubber stopper (17) in the injection tube (10); The support mechanism comprises a first connecting component and a second connecting component, wherein the first connecting component is detachably fixedly connected to a microscope condenser (1), a cell pipette (7) is detachably mounted on the first connecting component, and the angle of the cell pipette (7) on the first connecting component is adjustable; the second connecting component is detachably fixedly connected to an inverted microscope stage (14); and the injection tube (10) is fixedly mounted on the second connecting component.

2. A single cell clone picking device according to claim 1, characterized in that: The suction adjustment assembly comprises a first screw (19) and an adjustment knob (12), wherein the first screw (19) is threadedly connected to the threaded inner wall of the injection tube (10), and the two ends of the first screw (19) are respectively connected to the rubber stopper (17) and the adjustment knob (12), and the adjustment knob (12) is located outside the injection tube (10).

3. The single cell clone picking device according to claim 1, characterized in that: The cell pipette port (15) is arranged vertically.

4. The single cell clone picking device according to claim 1, characterized in that: The first connecting assembly comprises a first clamp (2), a second clamp (6), a lifting rod (5), a connecting seat (22) and two connecting pieces (3); the first clamp (2) is provided with a first clamping area for clamping a microscope condenser (1) and a second clamping area for clamping the connecting seat (22); the lifting rod (5) is mounted on the connecting seat (22); the second clamp (6) is mounted on the end of the lifting rod (5) via one of the connecting pieces (3); and the second clamp (6) is used to clamp a cell pipette (7).

5. The single cell clone picking device according to claim 4, characterized in that: The lifting rod (5) is inserted into a slot of the connecting seat (22) and is slidably connected to the connecting seat (22). A toothed portion is provided on one side of the lifting rod (5), and the toothed portion is meshed with a gear (21). The gear (21) is rotatably connected to the connecting seat (22).

6. The single cell clone picking device according to claim 4, characterized in that: The first clamp (2) comprises two metal sheets, one end of the two metal sheets being fixedly connected via a connector (3), and the other end of the two metal sheets clamping the microscope condenser (1) are then fixed to the connecting seat (22) via the connector (3).

7. The single cell clone picking device according to claim 6, characterized in that: The clamping surface of the metal sheet is provided with anti-slip and anti-collision rubber.

8. A single cell clone picking device according to any one of claims 4 to 7, characterized in that: The connecting member (3) comprises a connecting protrusion (31), a polished rod (32) and a threaded sleeve (34); one end of the polished rod (32) is connected to the connecting protrusion (31); the other end of the polished rod (32) is provided with a threaded portion (33); and the threaded portion (33) is threadedly connected to the threaded sleeve (34).

9. The single cell clone picking device according to claim 8, characterized in that: The second clamp (6) includes two metal sheets, between which is a third clamping area for clamping the cell pipette (7), and the two ends of the two metal sheets are fixed by a connector (3), one of the light rods (32) sequentially passes through one end of the two metal sheets and is threadedly connected to one of the threaded sleeves (34); the other light rod (32) sequentially passes through the other end of the two metal sheets and the lifting rod (5) and is threadedly connected to the other threaded sleeve (34).

10. The single cell clone picking device according to claim 1, characterized in that: The second connecting assembly comprises a connecting seat (11) and a fixing clamp (13), wherein the two ends of the connecting seat (11) are fixedly connected to the injection tube (10) and the fixing clamp (13), respectively, and the fixing clamp (13) is clamped and arranged on the inverted microscope stage (14).

Images