CN215655764U

CN215655764U - Automatic cell washing device

Info

Publication number: CN215655764U
Application number: CN202120534345.0U
Authority: CN
Inventors: 李青峰; 周双白; 高一鸣
Original assignee: Ninth Peoples Hospital Shanghai Jiaotong University School of Medicine
Current assignee: Ninth Peoples Hospital Shanghai Jiaotong University School of Medicine
Priority date: 2021-03-15
Filing date: 2021-03-15
Publication date: 2022-01-28
Anticipated expiration: 2031-03-15

Abstract

The utility model relates to the technical field of medical instruments, in particular to an automatic cell washing device, which comprises a sample adding unit and a centrifugal unit, wherein the sample adding unit comprises a first conduit and a sample adding device, and the first conduit is connected with the sample adding device; the centrifugal unit includes the centrifuge rotor of V-arrangement centrifuging tube and adaptation V-arrangement centrifuging tube, the application of sample ware is arranged in to the application of sample in the V-arrangement centrifuging tube. The automatic cell washing device provided by the utility model effectively removes impurities in the cell suspension through repeated washing and centrifugation of the V-shaped centrifuge tube. In the centrifugal process, under the action of centrifugal force, liquid is automatically balanced in the two arms of the V-shaped centrifugal tube without additionally arranging a balance tube. The process of liquid feeding and waste liquid discharging is full-automatic, repeated manual operation is reduced, and efficiency is improved.

Description

Automatic cell washing device

Technical Field

The utility model relates to the technical field of medical instruments, in particular to an automatic cell washing device.

Background

In the fields of medicine and cell biology, cell-related manipulations are often involved, including centrifugation and washing of cell suspensions. In the process of extracting primary cells from tissues or subculturing cells, since the components of the tissues or the culture medium are complicated, the extracted cell suspension contains many extracellular matrices other than the cellular components or impurity components other than the target cells, and these need to be removed. Because solid particles of different densities or particle sizes settle at different rates in a liquid, centrifuges are capable of separating liquids from solids using centrifugal forces. The centrifugal machine mainly comprises four parts, namely a driving part, a speed control part, a temperature control part, a vacuum system and a rotating head, and liquid with the same weight as that of the centrifugal machine to be centrifuged is required to be balanced before centrifugation.

The washing process of the cells comprises the steps of firstly adding cell suspension containing impurities into a centrifuge tube, placing the centrifuge tube in a centrifuge for centrifugation with certain centrifugal force and time, compared with the impurities in the suspension or PBS buffer solution for washing, allowing the cells with a slightly large sedimentation coefficient to sink to the bottom of the centrifuge tube under the action of centrifugal force and be attached to the wall of the centrifuge tube, removing the upper liquid containing the impurities, adding liquid for washing the cells such as PBS, blowing and beating the cells at the bottom of the centrifuge tube, suspending the cells in the buffer solution, uniformly distributing the cells in the liquid, and repeatedly centrifuging, adding liquid and suspending for 2-3 times to remove the impurity substances. The whole process of using the traditional centrifuge to wash cells needs repeated manual operation, consumes long time and is complex to operate, and the repeated pouring and transferring of liquid is easy to cause cell pollution.

SUMMERY OF THE UTILITY MODEL

In view of the above-mentioned disadvantages of the prior art, it is an object of the present invention to provide an automatic cell washing apparatus for solving the problems of the prior art.

In order to achieve the above and other related objects, the present invention provides an automatic cell washing apparatus, comprising a sample application unit and a centrifugation unit, wherein the sample application unit comprises a first conduit and a sample applicator, and the first conduit is connected to the sample applicator; the centrifugal unit includes the centrifuge rotor of V-arrangement centrifuging tube and adaptation V-arrangement centrifuging tube, the application of sample ware is arranged in to the application of sample in the V-arrangement centrifuging tube.

As described above, the automatic cell washing apparatus of the present invention has the following advantageous effects:

(1) through repeated washing and centrifugation of the V-shaped centrifuge tube, impurities in the cell suspension are effectively removed.

(2) In the centrifugal process, under the action of centrifugal force, liquid is automatically balanced in the two arms of the V-shaped centrifugal tube without additionally arranging a balance tube.

(3) The process of liquid feeding and waste liquid discharging is full-automatic, repeated manual operation is reduced, and efficiency is improved.

Drawings

FIG. 1 is a schematic view of an automatic cell washing apparatus according to the present invention.

FIG. 2 is another schematic view of the automatic cell washing apparatus of the present invention.

Description of the element reference numerals

1 sample adding unit

11 first conduit

111 liquid inlet conduit

112 cell entry catheter

113 first valve

12 sample injector

121 power device

122 sample adding head

123 sample adding needle

2 centrifugal Unit

21V-shaped centrifugal tube

Alpha inner included angle

211 first opening

212 second opening

Beta outer included angle

2121 second valve

213 Scale

22 centrifuge rotor

221 rotor barrel

2211 second conduit

2211a liquid conduit

2211b cell conduit

222 rotor cover

2221 channel

Detailed Description

The following description of the embodiments of the present invention is provided for illustrative purposes, and other advantages and effects of the present invention will become apparent to those skilled in the art from the present disclosure.

Please refer to fig. 1-2. It should be understood that the structures, ratios, sizes, and the like shown in the drawings and described in the specification are only used for matching with the disclosure of the specification, so as to be understood and read by those skilled in the art, and are not used to limit the conditions under which the present invention can be implemented, so that the present invention has no technical significance, and any structural modification, ratio relationship change, or size adjustment should still fall within the scope of the present invention without affecting the efficacy and the achievable purpose of the present invention. In addition, the terms "upper", "lower", "left", "right", "middle" and "one" used in the present specification are for clarity of description, and are not intended to limit the scope of the present invention, and the relative relationship between the terms and the terms is not to be construed as a scope of the present invention.

As shown in the figure, the present invention provides an automatic cell washing device, which includes a sample adding unit 1 and a centrifugation unit 2, wherein the sample adding unit 1 includes a first conduit 11 and a sample adding device 12, and the first conduit 11 is connected to the sample adding device 12; the centrifuge unit 2 comprises a V-shaped centrifuge tube 21 and a centrifuge rotor 22 adapted to the V-shaped centrifuge tube 21, and the sample applicator 12 is used for applying samples to the V-shaped centrifuge tube 21.

The first conduit 11 is used to add and transfer cells or wash solution into the automatic cell washing apparatus. The first conduit 11 is provided with a first valve 113. In one embodiment, the first valve 113 is a three-way valve. Preferably, the switch is an electric control three-way switch.

The first conduit 11 is provided with one or more. In one embodiment, the first conduit 11 is provided with two, respectively, an inlet conduit 111 and a cell inlet conduit 112. The liquid inlet pipe 111 is used for feeding and transporting the washing liquid, and the cells enter the pipe 112 for feeding and transporting the cells to be washed. The first conduit C11 is a flexible conduit, and provides a space for the sample addition head C122 to move.

The sample injector 12 includes a power device 121, a sample injection head 122 and a sample injection needle 123, the power device 121 is electrically connected to the sample injection head 122, one end of the sample injection head 122 is connected to the first conduit 11, and the other end is connected to the sample injection needle 123.

The power device 121 drives the sample adding head 122 to further drive the sample adding needle 123 to add sample to the V-shaped centrifuge tube 21. In one embodiment, the motive device 121 is selected from a linear motor. The power unit C121 is fixed and the sample addition head C122 moves on its guide rail.

In one embodiment, the sample addition head 122 is a holder for the sample addition needle 123, so that the sample addition needle 123 can move up and down along the guide rail thereof along with the power device 121.

In one embodiment, the sample addition needle 123 is a hollow syringe-like needle.

The cross section of the V-shaped centrifuge tube 21 is V-shaped. The angle of the inner included angle α of the V-shape is not limited.

A first opening 211 is arranged at the internal included angle alpha of the V-shaped centrifuge tube 21. The first opening 211 may be used to add a sample to the V-shaped centrifuge tube 21.

A one-way channel is arranged at the first opening 211. In one embodiment, the one-way channel is a rubber stopper. The sample adding needle 123 moves downwards to puncture the rubber stopper and add the sample into the V-shaped centrifuge tube 21. The sample adding needle 123 withdraws the rubber stopper when moving upwards, and the rubber stopper is sealed again to prevent the liquid from flowing out.

The one-way passage may be replaced with a one-way valve. The application of sample needle 123 moves down, can carry out application of sample operation behind the back-open check valve, when application of sample needle 123 moves up and withdraws from, then the valve is closed, prevents that liquid from flowing out.

And a second opening 212 is arranged at the external included angle beta of the V-shaped centrifuge tube 21. The second opening 212 may be used to add or remove samples from the V-shaped centrifuge tube 21.

In one embodiment, a second valve 2121 is disposed at the second opening 212. The second valve 2121 is used to open or close the second opening 212. In one embodiment, the second valve 2121 is also used to determine which conduit the liquid flows from. In one embodiment, the second valve 2121 is a two-way switch or a three-way switch. The second valve 2121 is an electric control switch or a manual switch. When the electric control switch is set, the automatic cell washing device is used in combination with a controller, and the controller is electrically connected with the second valve 2121, so that the second valve 2121 is controlled, and the purpose of controlling liquid to flow out from different catheters is achieved. When a manual switch is provided, the V-tube 21 can be removed from the centrifuge when liquid discharge is required, and the second valve 2121 is manually controlled to select an appropriate tube to discharge the liquid.

The two arms of the V-shaped centrifuge tube 21 are respectively cylindrical, and the tail part of the cylinder is conical or hemispherical.

The wall of the V-shaped centrifuge tube 21 is provided with scales 213. The scale 213 is used to measure the volume of liquid in the centrifuge tube. The scale 213 can be set at different positions on the tube wall or set as a zero scale according to the specific use condition of the V-shaped centrifuge tube 21. For example, when the first opening 211 is used as an inlet of a centrifuge tube, since the liquid is first concentrated at the second opening 212 after entering, the zero scale can be set near the second opening 212; or when the second opening 212 is used as the inlet of the centrifuge tube, the liquid is firstly concentrated at the tail parts of the two arms of the centrifuge tube after entering, and the zero scale can be arranged at the tail part at the moment.

The centrifuge rotor 22 comprises a rotor barrel 221 and a rotor cover 222 which are matched with each other, the rotor barrel 221 is provided with a V-shaped inner groove, the rotor cover 222 is provided with a conical part, and the V-shaped inner groove and the conical part are matched with the V-shaped centrifuge tube 21 in shape. The bottom of the V-shaped inner groove is provided with a concave part. The recess is for placement of a second valve 2121.

In one embodiment, the rotor cover 222 has a passageway 2221 on its central axis. The central shaft is as follows: in the working state, the vertical line made from the vertex of the conical part to the horizontal plane is crossed. The channel 2221 is used for loading the sample pin 123 through it into the V-shaped centrifuge tube 21.

A second conduit 2211 is arranged inside the rotor barrel 221, and an opening at one end of the second conduit 2211 is arranged at the bottom of the V-shaped inner groove. When the V-shaped centrifuge tube 21 is placed in the V-shaped inner trough, the second opening 212 is connected to the opening of the second conduit 2211 via the second valve 2121. The second conduit 2211 is provided with one or more. When a second conduit 2211 is provided, cells and fluid enter or exit the same conduit. In one embodiment, the second conduit 2211 is provided with two, a liquid conduit 2211a and a cell conduit 2211 b.

The end of the second conduit 2211 is also provided with a recovery bottle. A common reagent bottle may be used as the recovery bottle. The recovery bottle comprises a cell recovery bottle and a waste liquid recovery bottle.

The bottom of the rotor barrel 221 is provided with a rotor rotating shaft. In one embodiment, the rotor shaft is hollow.

The automatic cell washing device needs to be arranged in a centrifuge to be matched with the centrifuge for use. The type and model of the centrifuge are not particularly limited. The automatic cell washing device may also be used in conjunction with a fat treatment device.

The automatic cell washing device can be used to wash various cells, such as adipocytes.

One method of using the automatic cell washing device is as follows: install V-arrangement centrifuging tube 21 in centrifuge rotor 22, install centrifuge rotor 22 in centrifuge, sample application needle 123 is fixed in on sample application head 122, linear motor drive sample application head 122, further drive sample application needle 123 downstream, sample application needle 123 passes the check valve, open first valve 113, it adds V-arrangement centrifuging tube 21 with the cell to get into first pipe 11, the liquid volume can be observed through the scale on V-arrangement centrifuging tube 21's the pipe wall, make the liquid volume be in the scope that is most suitable to handle, the liquid level is between the highest and the minimum scale of V-arrangement centrifuging tube 21. The sampling needle 123 is pulled out, and the check valve is closed to prevent the liquid from flowing out. The centrifugal time and the centrifugal force are set on the centrifugal machine or other control equipment, the centrifugation is started according to the preset centrifugal time and the preset centrifugal force, the liquid flows to the two arms of the V-shaped centrifugal tube 21 under the action of the centrifugal force, and the liquid volume distribution of the two arms is uniform. After centrifugation, slowly stop rotatory, the cell adheres to the afterbody of V-arrangement centrifuging tube 21 both arms under the effect of centrifugal force, and liquid slowly assembles second opening 212 department again, opens second valve 2121 and discharges the waste liquid, and the waste liquid flows out and is collected in order to prevent that liquid splashes along second pipe 2211 by the waste liquid collecting bottle, closes second valve 2121 after the waste liquid discharges. The first valve 113 is opened, PBS is rapidly added through the liquid adding conduit, the first valve 113 is automatically closed after the liquid level of the added liquid reaches a proper water level line, the linear motor is raised again, the liquid adding needle 123 is withdrawn, the rotor rapidly rotates to drive the V-shaped centrifuge tube 21, adhered cells are washed off, the cells are prevented from being dried, and the cells and the PBS are mixed for heavy suspension. After resuspension, centrifugation is performed again, the cells are attached to the top of the two arms of the V-shaped centrifuge tube 21 again, the second valve 2121 is opened, and waste fluid is discharged from the second conduit 2211, and the second valve 2121 is closed after waste fluid. Repeating the above washing process for 2-3 times. And finally, in washing, after PBS is added, the rotor is quickly started to rotate for 2-3 times to drive the V-shaped centrifuge tube 21 to rotate, the PBS quickly washes the cells adhered to the top of the V-shaped centrifuge tube 21 from the bottom to the top of the V-shaped centrifuge tube 21 to prevent the cells from being dried, and the cells and the PBS are mixed and resuspended. The cell-containing PBS cell suspension is allowed to re-collect under gravity to the second opening 212. The second valve 2121 is opened to discharge the cells from the second conduit 2211. Alternatively, after the last resuspension of the cells, the V-tube 21 is removed from the centrifuge rotor 22 and the second valve 2121 is manually opened to collect the cell suspension.

In conclusion, the present invention effectively overcomes various disadvantages of the prior art and has high industrial utilization value.

The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the utility model. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.

Claims

1. An automatic cell washing device, characterized in that the automatic cell washing device comprises a sample adding unit (1) and a centrifugation unit (2), the sample adding unit (1) comprises a first conduit (11) and a sample adding device (12), the first conduit (11) is connected with the sample adding device (12); the centrifugal unit (2) comprises a V-shaped centrifugal tube (21) and a centrifuge rotor (22) matched with the V-shaped centrifugal tube (21), and the sample adding device (12) is used for adding samples into the V-shaped centrifugal tube (21).

2. The automatic cell washing device according to claim 1, wherein the first conduit (11) is provided with a first valve (113).

3. An automatic cell washing device according to claim 1, characterized in that said first conduit (11) is provided with one or more.

4. The automatic cell washing device according to claim 1, wherein the first conduit (11) is provided with two, respectively, a liquid inlet conduit (111) and a cell inlet conduit (112).

5. The automatic cell washing device according to claim 1, wherein the sample applicator (12) comprises a power device (121), a sample application head (122), and a sample application needle (123), wherein the power device (121) is electrically connected to the sample application head (122), and wherein one end of the sample application head (122) is connected to the first conduit (11) and the other end is connected to the sample application needle (123).

6. The automated cell washing apparatus of claim 1, wherein the V-shaped centrifuge tube (21) further comprises one or more of the following features:

1) a first opening (211) is arranged at the inner included angle (alpha);

2) a second opening (212) is arranged at the outer included angle (beta);

3) the pipe wall is provided with scales (213).

7. The automatic cell washing device according to claim 6, wherein a one-way valve is provided at the first opening (211); or a second valve (2121) is arranged at the second opening (212).

8. The automatic cell washing device of claim 1, wherein the centrifuge rotor (22) comprises a rotor barrel (221) and a rotor cover (222) adapted to each other, the rotor barrel (221) being provided with a V-shaped inner groove and the rotor cover (222) being provided with a tapered portion, both the V-shaped inner groove and the tapered portion matching the shape of the V-shaped centrifuge tube (21).

9. The automatic cell washing device according to claim 1, wherein a passage (2221) is provided on a central shaft of the rotor cover (222).

10. The automatic cell washing device according to claim 8, wherein the rotor barrel (221) is provided with a second conduit (2211) inside, an opening at one end of the second conduit (2211) is arranged at the bottom of the V-shaped inner groove, and the second opening (212) is connected with the opening at one end of the second conduit (2211) after the V-shaped centrifuge tube (21) is arranged in the V-shaped inner groove.

11. The automatic cell washing device according to claim 10, wherein the second conduit (2211) is provided with one or more.

12. The automatic cell washing device according to claim 10, wherein the second conduit (2211) is provided with two, a liquid conduit (2211a) and a cell conduit (2211b), respectively.

13. The automatic cell washing device according to claim 10, wherein the second conduit (2211) is further provided with a recovery bottle at the end.

14. The automated cell washing apparatus of claim 13, wherein the recovery vial comprises a cell recovery vial and a waste fluid recovery vial.