CN111004717A

CN111004717A - Single cell separation and collection device

Info

Publication number: CN111004717A
Application number: CN201911362924.5A
Authority: CN
Inventors: 刘迁; 金建华; 陆文斌; 杨春霞; 李文晶; 沈颖
Original assignee: Changzhou Wujin Peoples Hospital
Current assignee: Changzhou Wujin Peoples Hospital
Priority date: 2019-12-26
Filing date: 2019-12-26
Publication date: 2020-04-14

Abstract

The invention relates to the technical field of medical instruments, in particular to a single cell separating and collecting device, which comprises a microscope, wherein a culture dish is placed on an object stage of the microscope, a support is fixed on a frame of the microscope, a suction pipe assembly is installed on the support and comprises a suction pipe, the suction pipe extends into the culture dish to suck single cells, the tail end of the suction pipe assembly is communicated with a liquid suction pipeline, and a suction device is arranged in the liquid suction pipeline.

Description

Single cell separation and collection device

Technical Field

The invention relates to the technical field of medical instruments, in particular to a single cell separating and collecting device.

Background

In natural science and basic medicine, immune cells or adherent tumor cells which are interesting or specific are selected from primary cultured cells or solid tumor cells, or the special cells which are interesting are separated, cultured or detected after transfection and drug screening. At present, import equipment (such as a flow sorter, a micropore chip separator and the like) commonly used in laboratories for separating cells is expensive, and related equipment can only separate a certain percentage of specific cells, wherein the specific cells containing impurities cannot be removed; the technique of using manually separated aspiration tubes requires a high level of operator skill, and the micromanipulation technique is currently directed to yeast cells. Therefore, an instrument which can be used for culturing or detecting after separating single cells provides great convenience for researchers or medical technicians engaged in cell culture at present. .

Disclosure of Invention

The invention provides a single cell separating and collecting device which is convenient for absorbing single cells, and aims to solve the problems that impurity cells existing in a high-throughput sorting instrument in the prior art cannot be completely removed, the instrument is expensive, single cells are not stably absorbed manually, and the like.

The technical scheme adopted by the invention for solving the technical problems is as follows:

the utility model provides a single cell separation and collection device, includes microscope, separator and collection device, microscope's objective table on placed the culture dish, separator include the support, the support on install the straw subassembly, the straw subassembly include the straw, the straw stretches and absorbs single cell in the culture dish, collection device including the intercommunication at the straw way of sucking of straw subassembly tail end, the straw way in be provided with suction means.

Furthermore, the suction device comprises a fluid pump arranged in the liquid suction pipeline, a purification branch is arranged at the upstream of the fluid pump in the liquid suction pipeline, sterile air filter cotton is arranged on the purification branch, and a stop valve is further arranged on the purification branch.

Furthermore, the liquid suction pipeline in be equipped with first branch and second branch, first branch is close to the liquid suction pipeline import, the second branch is close to the liquid suction pipeline export, suction means is including setting up the blower pump on first branch and setting up the aspirator pump on the second branch, all is equipped with the stop valve on first branch and the second branch, be equipped with aseptic air filter cotton on the first branch.

Further, the straw assembly still include installation pipe and base, the base fix the tip at the installation pipe, the installation intraductal flexible spacer bush that is equipped with, the straw parcel in flexible spacer bush, and the straw front end stretches into in the culture dish.

Furthermore, the support is fixed beside the microscope, the support comprises an upright post and a cross beam arranged on the upright post, a height adjusting device is arranged between the upright post and the cross beam, and the mounting pipe is rotatably connected to the end part of the cross beam.

Furthermore, the support is fixed on a frame of the microscope, the installation pipe is rotatably connected to the support, and the installation pipe rotates along the direction close to the objective table and the direction far away from the objective table.

Furthermore, the support on rotate and connect the jack catch, the installation pipe press from both sides in the jack catch, the support still have the fixed knot who is used for fixing the jack catch rotational position to construct.

Furthermore, the fixing structure is a fixing pin or a buckle.

Furthermore, the flexible spacer sleeve is made of silica gel or foam, and the liquid suction pipeline is a transparent hose.

Further, the microscope and the separation device are covered with a protective cover for isolating bacteria and dust.

Has the advantages that:

(1) the single cell separating and collecting device provided by the invention replaces manpower to absorb single cells, the absorption process is stable, and the efficiency is high;

(2) the single cell separating and collecting device has simple structure, easy realization and low cost, and is convenient for popularization in a laboratory;

(3) the single cell separating and collecting device has certain accommodating capacity and can separate different types of cells simultaneously;

(4) the straw assembly is installed through the jack catch, can dismantle or change according to different specification demands, and application scope is wide.

Drawings

The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.

FIG. 1 is a schematic structural view of an installation tube and a straw assembly;

FIG. 2 is a schematic view showing the structure of the single cell separating and collecting apparatus according to example 1;

FIG. 3 is a schematic structural view of embodiment 2 of the single cell separating and collecting apparatus;

FIG. 4 is a view showing a structural example of the jaw;

fig. 5 is a schematic view showing an operating state of the transparent hose.

The device comprises a microscope 1, a microscope 2, a culture dish 3, a support 31, a column 32, a beam 4, a fluid pump 5, sterile filter cotton 6, a stop valve 7, a straw assembly 71, a straw 72, a base 73, an installation pipe 8, a flexible spacer sleeve 9, a jaw 10, a transparent hose 11, a purification branch 12, a first branch 13, a second branch 14, an air blowing pump 15 and an air suction pump.

Detailed Description

The utility model provides a single cell separation and collection device, includes microscope 1, separator and collection device, has placed culture dish 2 on microscope 1's the objective table, and separator includes support 3, installs straw subassembly 7 on the support 3, and straw subassembly 7 includes straw 71, and straw 71 stretches and absorbs single cell in culture dish 2, and collection device is provided with suction means including the intercommunication in the liquid suction pipeline of straw subassembly 7 tail end in the liquid suction pipeline. The microscope 1 and the separating apparatus are jointly externally covered with a protective cover for protecting against bacteria and dust. Mainly cover 2 positions of culture dish, the outside bacterium can be completely cut off to the protection casing, dust etc. expose outside such as the knob of microscope 1, the operation of being convenient for, moreover, transparent hose 10 also has been drawn forth, and the protection casing can be made detachable, perhaps opens the door in one side of support 3, the rotatory installation pipe 6 of being convenient for like this or put into culture dish 2 etc..

As shown in fig. 1, the straw assembly 7 further includes a mounting tube 6 and a base 72, the base 72 is fixed at an end of the mounting tube 6, a flexible spacer 8 is disposed in the mounting tube 6, the straw 71 is wrapped in the flexible spacer 8, and a front end of the straw 71 extends into the culture dish 2. The flexible spacer sleeve 8 is made of silica gel or foam, and the liquid absorption pipeline is a transparent hose 10. The transparent hose 10 is connected with the base 72, and the connection can be performed by adopting a quick connection mode. The shape of the pipette path in the figure is only a schematic diagram, and the actual shape or length is set according to the actual situation.

The suction device comprises the following embodiments:

example 1:

referring to fig. 2, the bracket 3 is fixed beside the microscope 1, the bracket 3 comprises a column 31 and a beam 32 mounted on the column, a height adjusting device is arranged between the column 31 and the beam 32, and the mounting tube 6 is rotatably connected to the end of the beam 32. The mounting tube 6 can be rotated to a position away from the stage of the microscope 1, so that the work of placing the culture dish 2 or cleaning the stage of the microscope 1 can be performed without collision interference with the straw assembly 7.

The liquid suction pipeline is internally provided with a first branch 12 and a second branch 13, the first branch 12 is close to the liquid suction pipeline inlet, the second branch 13 is close to the liquid suction pipeline outlet, the suction device comprises a blowing pump 14 arranged on the first branch 12 and a suction pump 15 arranged on the second branch 13, the first branch 12 and the second branch 13 are both provided with a stop valve 6, and the first branch 12 is provided with sterile air filter cotton 5.

Example 1 working principle:

before the suction is started, the suction tube 71 is blocked, then the stop valves 6 of the first branch 12 and the second branch 13 are opened, the blowing pump 14 and the suction pump are opened, the air is filtered through the sterile air filter cotton 5 in the first branch 12, and then the suction of the cells is started.

The process of cell suction is as follows: the left and right positions and the height of the culture dish 2 are adjusted by an adjusting knob of the microscope 1, then the required single cell is screened, the suction inlet of the suction pipe 71 is positioned below the microscope 1, when the required single cell is found, the suction can be carried out, firstly, the stop valve 6 and the air blowing pump 14 of the first branch are closed, the stop valve 6 and the air sucking pump 15 of the second branch are opened, the cell is sucked, the transparent hose 10 between the first branch 12 and the second branch 13 is relatively longer, then, the sucked cells are positioned in the transparent hose 10 between the first branch 12 and the second branch 13, the suction can be carried out after a period of time, and thus, as shown in fig. 5, different single cells can be conveyed in the transparent hose 10 at intervals. In FIG. 5, gray represents a liquid portion, white represents air, and small black dots represent single cells, and after the completion of the aspiration, the cut-off valve 6 of the first branch and the blower pump 14 are opened, and the cut-off valve 6 of the second branch and the aspirator pump 15 are closed, so that the cells are finally discharged to different containers such as a collector, etc. correspondingly under the action of the blower pump 14.

Example 2:

referring to fig. 3-4, the bracket 3 is fixed on the frame of the microscope, the mounting tube 6 is rotatably connected to the bracket 3, and the mounting tube 6 rotates in the direction of approaching and departing from the stage.

The suction device comprises a fluid pump 4 arranged in a liquid suction pipeline, a purification branch 11 is arranged at the upstream of the fluid pump 4 in the liquid suction pipeline, sterile air filter cotton 5 is arranged on the purification branch 11, and a stop valve 6 is further arranged on the purification branch 11.

The support 3 is rotatably connected with a clamping jaw 9, the mounting tube 6 is clamped in the clamping jaw 9, and the support 3 is also provided with a fixing structure for fixing the rotating position of the clamping jaw 9. The fixing structure is a fixing pin or a buckle.

Rotate on the support 3 and connect jack catch 9, installation pipe 6 presss from both sides in jack catch 9, during the installation, as long as open jack catch 9 one side screw, pack into installation pipe 6 after screw up again can, support 3 still is used for fixing the fixed knot who lives jack catch 9 turned position constructs. The fixing structure is a fixing pin or a buckle. In actual setting, the position of the installation tube 6 can be set to two grades, one grade is the working state position, the other grade is the roll-out position, and after the roll-out, the culture dish 2 can be placed or the microscope 1 can be cleaned, and the like, without collision and interference with the straw assembly 7.

Example 2 working principle:

before the cell suction is started, the suction pipe 71 is blocked, the stop valve 6 on the purification branch 11 is opened, the fluid pump 4 is opened to suck air, the air is filtered through the sterile air filter cotton 5, and the stop valve 6 is closed to start the cell suction.

The left and right positions and the height of the culture dish 2 are adjusted by the adjusting knob of the microscope 1, then the required single cell is screened, the suction inlet of the suction pipe 71 is positioned below the microscope 1, and when the required single cell is found, the fluid pump 4 can be driven to suck. The fluid pump 4 can be adjusted manually, for example, to suck 2 μ l of liquid each time, and then suck at intervals, so that different single cells are fed into the transparent tube 10 at intervals and finally discharged into different containers such as a collector, as shown in fig. 5, where gray represents the liquid portion, white represents air, and small black dots represent single cells.

It should be understood that the above-described specific embodiments are merely illustrative of the present invention and are not intended to limit the present invention. Obvious variations or modifications which are within the spirit of the invention are possible within the scope of the invention.

Claims

1. The single cell separating and collecting device is characterized by comprising a microscope (1), a separating device and a collecting device, wherein a culture dish (2) is placed on an object stage of the microscope (1), the separating device comprises a support (3), a sucker component (7) is installed on the support (3), the sucker component (7) comprises a needle tube (71), the needle tube (71) extends into the culture dish (2) to suck single cells, the collecting device comprises a sucker pipeline communicated with the tail end of the sucker component (7), and a sucking device is arranged in the sucker pipeline.

2. The apparatus for separating and collecting single cells according to claim 1, wherein: the suction device comprises a fluid pump (4) arranged in a liquid suction pipeline, a purification branch (11) is arranged at the upstream of the fluid pump (4) in the liquid suction pipeline, sterile air filter cotton (5) is arranged on the purification branch (11), and a stop valve (6) is further arranged on the purification branch (11).

3. The apparatus for separating and collecting single cells according to claim 1, wherein: the liquid suction pipeline is provided with a first branch (12) and a second branch (13), the first branch (12) is close to a liquid suction pipeline inlet, the second branch (13) is close to a liquid suction pipeline outlet, the suction device comprises a blowing pump (14) arranged on the first branch (12) and a suction pump (15) arranged on the second branch (13), stop valves (6) are arranged on the first branch (12) and the second branch (13), and sterile air filter cotton (5) is arranged on the first branch (12).

4. The apparatus for separating and collecting single cells according to claim 1, wherein: the straw assembly (7) still include installation pipe (6) and base (72), base (72) fix the tip at installation pipe (6), installation pipe (6) in be equipped with flexible spacer (8), straw (71) parcel in flexible spacer (8), and straw (71) front end stretch into in culture dish (2).

5. The apparatus for separating and collecting single cells according to claim 4, wherein: the flexible spacer bush (8) is made of silica gel or foam, and the liquid suction pipeline is a transparent hose (10).

6. The apparatus for separating and collecting single cells according to claim 4, wherein: the microscope is characterized in that the support (3) is fixed beside the microscope (1), the support (3) comprises a stand column (31) and a cross beam (32) installed on the stand column, a height adjusting device is arranged between the stand column (31) and the cross beam (32), and the installation tube (6) is rotatably connected to the end part of the cross beam (32).

7. The apparatus for separating and collecting single cells according to claim 4, wherein: support (3) fix in microscope's frame, installation pipe (6) rotate to be connected support (3) on, installation pipe (6) rotate along the direction of being close to the objective table and keeping away from the objective table.

8. The single cell separation and collection device according to claim 6 or 7, wherein: support (3) on rotate and connect jack catch (9), installation pipe (6) press from both sides in jack catch (9), support (3) fixed knot who is used for fixing jack catch (9) turned position constructs in addition.

9. The apparatus for separating and collecting single cells according to claim 8, wherein: the fixing structure is a fixing pin or a buckle.

10. The apparatus for separating and collecting single cells according to claim 1, wherein: the microscope (1) and the separating device are covered with a protective cover for isolating bacteria and dust.