CN211179220U

CN211179220U - A device that is used for mesenchymal stem cell surface marker immunochemifluorescence to dye

Info

Publication number: CN211179220U
Application number: CN201922236236.6U
Authority: CN
Inventors: 牟丽莎; 陆赢; 蒲祖辉; 赵萌; 娄琪
Original assignee: Shenzhen Lansi Artificial Intelligence Medical Research Institute; Shenzhen Second Peoples Hospital
Current assignee: Shenzhen Lansi Artificial Intelligence Medical Research Institute; Shenzhen Second Peoples Hospital
Priority date: 2019-12-13
Filing date: 2019-12-13
Publication date: 2020-08-04
Anticipated expiration: 2029-12-13

Abstract

The application discloses a device for immunochemical fluorescent staining of mesenchymal stem cell surface markers. The device comprises a culture dish main body and a culture tank side wall; the side wall of the culture tank is arranged in the accommodating cavity of the culture dish main body, the side wall of the culture tank surrounds the accommodating cavity of the culture dish main body to form an independent culture tank, and the side wall of the culture tank is in a horn-shaped design with an upward opening; the side wall of the culture tank is provided with a first through hole and a second through hole which can be opened and closed at the symmetrical positions of the bottom of the side wall; the bottom of the side wall of the culture dish main body is provided with a culture dish through hole capable of being opened and closed. The device can simply and conveniently replace the culture solution or the chemical fluorescent dye, and improves the quality and the efficiency of the mesenchymal stem cell immunochemical fluorescent staining; and the using amount of a culture medium and an antibody can be reduced, and the waste is reduced, so that the detection cost of the immunochemical fluorescent staining of the mesenchymal stem cell surface marker is reduced.

Description

A device that is used for mesenchymal stem cell surface marker immunochemifluorescence to dye

Technical Field

The application relates to the field of mesenchymal stem cell detection devices, in particular to a device for immunochemical fluorescent staining of mesenchymal stem cell surface markers.

Background

The heterogeneity of Mesenchymal Stem Cells (MSCs), i.e. the subset that supports hematopoietic maintenance hematopoietic stem cell (abbreviated HSC) function to enhance engraftment efficiency, the paracrine angiogenic subset, and the subset that immunoregulation suppresses the development of GVDH, has not been well defined. This is a major problem that causes current MSC therapy to face clinically unstable therapeutic effects.

Recent studies found that a series of functional markers of MSC, such as Nestin-GFP labeled bone marrow MSC cells and HSC form a common microenvironment, PDGFR α and CD5115 positive MSC cells can promote HSC proliferation, L epr expressed MSC is a key cell of bone marrow osteogenesis, Mx1-Cre can be labeled to MSC cells with high osteogenic capacity in bone marrow, Gremlin1 expressed cells have multipotent differentiation potential of osteogenic, chondrogenic and reticular stromal cells, and Gli1 positive perivascular MSC cells can promote repair after injury of various tissues.

Therefore, the identification of different subpopulations of MSCs is very important. The immunochemical (fluorescent) staining identification of MSCs with different surface markers is usually performed by culturing cells on circular cover slips in a cell culture plate, fixing the MSCs for chemofluorescent staining after the MSCs are cultured, and adding or removing liquids such as antibodies, eluents and the like to or from the culture plate by using a pipettor for many times during the staining process. Because the process involves multiple steps, the number of MSCs on the slide is often reduced and the cell loss during the washing process is high; meanwhile, when the chemical fluorescent staining operation is performed, the sheet is often dried due to antibody loss, or a large amount of antibody must be used, causing unnecessary loss. The steps of the whole experimental process are complicated, and the repeatability and the efficiency of the experiment are not high. Therefore, improving the quality and efficiency of MSC immunochemical fluorescent staining has also been one of the important contents of MSC identification.

Disclosure of Invention

The purpose of this application provides an improved device that is used for mesenchymal stem cell surface marker immunochemical fluorescence staining.

In order to achieve the purpose, the following technical scheme is adopted in the application:

the application discloses a device for immunochemical fluorescent staining of mesenchymal stem cell surface markers, which comprises a culture dish main body and a culture tank side wall; the side wall of the culture tank is arranged in the accommodating cavity of the culture dish main body, the side wall of the culture tank surrounds the accommodating cavity of the culture dish main body to form an independent culture tank, and the side wall of the culture tank is in a flared design with an upward opening, so that the culture tank is in an inverted cone frustum shape; the side wall of the culture tank is provided with a first through hole and a second through hole which can be opened and closed at the symmetrical positions of the bottom of the side wall; the bottom of the side wall of the culture dish main body is provided with a culture dish through hole capable of being opened and closed.

In the device, a horn-shaped culture tank is designed in the culture dish, and the horn-shaped culture tank meets the space required by cell culture growth on one hand and can reduce the use amount of culture medium and antibodies on the other hand. In addition, when cleaning is carried out, the first through hole and the second through hole which can be opened and closed are opened, the culture tank is communicated with the accommodating cavity of the culture dish, more liquid can be used for elution, and the antibody can be eluted better; after the elution is finished, the washing liquid can be poured out by directly opening the through hole of the culture dish. The device of this application directly carries out the change of chemi-fluorescent dye or elution liquid through first through-hole, second through-hole and culture dish through-hole, and convenience simple to use has improved MSC immunochemistry fluorescence staining's quality and efficiency.

It should be noted that, the openable design of the first through hole and the second through hole, and the openable design of the culture dish through hole may refer to the existing opening and closing design of the through hole or the pipe. In an implementation of this application, first through-hole and second through-hole adopt baffle control to open and shut, and the culture dish through-hole adopts piston control to open and shut.

Therefore, preferably, the apparatus of the present application further comprises a first baffle and a second baffle; the first baffle plate can be arranged on the side wall of the culture tank in a vertically sliding manner and is arranged at the position of the first through hole; the first through hole is opened or closed by moving the first baffle up and down; the second baffle plate can be arranged on the side wall of the culture tank in a vertically sliding manner and is arranged at the position of the second through hole; the opening or closing of the second through hole is realized by moving the second baffle plate up and down.

It should be noted that, the first baffle and the second baffle are installed in a vertically sliding manner, and may be clamped by a slide rail or in other conventional manners, which is not limited herein.

Preferably, the device of the present application further comprises a piston, the piston is installed in the culture dish through hole, and the culture dish through hole is closed or opened by plugging or pulling the piston.

It should be noted that the effect of piston is that the jam or open the culture dish through-hole, therefore, in an implementation of this application, specifically adopt the plug, the interference fit through the plug realizes that the closely of culture dish through-hole closes.

Preferably, the culture trough is located at the center of the accommodating cavity of the culture dish body, and the center of the culture trough overlaps with the center of the accommodating cavity of the culture dish body.

Preferably, the chamber that holds of culture dish main part is cylindrical, and the culture tank is the inverted cone frustum shape by the culture tank lateral wall encloses to close, and the axis that holds the chamber of culture dish main part coincides with the axis of the culture tank of cone frustum shape.

Preferably, the culture dish body and the side walls of the culture tank are transparent. For example, transparent glass or other transparent plastic material is used to make the transparent.

Due to the adoption of the technical scheme, the beneficial effects of the application are as follows:

the device for the immunochemical fluorescent staining of the marker on the surface of the mesenchymal stem cell not only can simply and conveniently replace the culture solution or the chemical fluorescent dye, but also can improve the quality and the efficiency of the immunochemical fluorescent staining of the mesenchymal stem cell; and the using amount of a culture medium and an antibody can be reduced, and the waste is reduced, so that the detection cost of the immunochemical fluorescent staining of the mesenchymal stem cell surface marker is reduced.

Drawings

Fig. 1 is a sectional view of an apparatus for immunochemical fluorescent staining of mesenchymal stem cell surface markers in an embodiment of the present application;

fig. 2 is a sectional view of another use state of the device for the immunochemical fluorescent staining of mesenchymal stem cell surface markers in the embodiment of the present application.

Detailed Description

This application is used for device of mesenchymal stem cell surface marker immunochemistry fluorescence staining improves on the basis of current culture dish and forms, and specific the increase sets up the tubaeform culture tank that encloses by the culture tank lateral wall in the culture dish inside holds the intracavity to realize the required chemifluorescence staining function of this application, and, can reduce the use amount of culture medium and antibody, convenience simple to use has improved MSC immunochemistry fluorescence staining's quality and efficiency.

The present invention will be described in further detail with reference to the following detailed description and accompanying drawings. The following examples are intended to be illustrative of the present application only and should not be construed as limiting the present application.

Examples

The device for the immunochemical fluorescent staining of the mesenchymal stem cell surface marker of the present example, as shown in fig. 1 and 2, comprises a culture dish body 1, a culture tank side wall 2, a first baffle 3, a second baffle 4 and a piston 5. The culture tank side wall 2 is arranged in the accommodating cavity of the culture dish main body 1, the culture tank side wall 2 encloses the accommodating cavity of the culture dish main body 1 to form an independent culture tank 20, and the culture tank side wall 2 is in a horn-shaped design with an upward opening, so that the culture tank is in an inverted frustum shape, as shown in fig. 1; the bottom of the side wall 2 of the culture tank is provided with a first through hole 21 and a second through hole 22 which can be opened and closed at symmetrical positions; the bottom of the side wall of the culture dish main body 1 is provided with a culture dish through hole 11 which can be opened and closed. In one implementation manner of this embodiment, the first through hole 21 and the second through hole 22 can be opened and closed by installing the first baffle 3 on the side wall 2 of the culture tank in a manner of sliding up and down, and installing the baffle at the position of the first through hole 21; the opening or closing of the first through hole 21 is achieved by moving the first shutter 3 up and down. Similarly, the second baffle 4 is mounted on the side wall 2 of the culture tank in a manner of sliding up and down and is mounted at the position of the second through hole 22; the opening or closing of the second through hole 22 is achieved by moving the second shutter 4 up and down. In one implementation manner of this embodiment, the openable and closable manner of the culture dish through hole 11 is that the piston 5 is installed in the culture dish through hole 11, and the culture dish through hole 11 is closed or opened by plugging or unplugging the piston 5. The accommodating chamber of the culture dish main body 1 of the present embodiment is cylindrical, the culture tank 20 is enclosed by the culture tank side wall 2 to form an inverted truncated cone shape, and the central axis of the accommodating chamber of the culture dish main body 1 coincides with the central axis of the truncated cone-shaped culture tank 20, that is, the culture tank 20 is disposed at the central position of the accommodating chamber of the culture dish main body 1. The device for the immunochemical fluorescent staining of the mesenchymal stem cell surface marker is transparent.

When the elution liquid needs to be replaced, the piston 5 is pulled out, and the piston 5 is restored after the liquid is replaced. When cell culture and antibody incubation are carried out, the first baffle plate 3 and the second baffle plate 4 are slid downwards, so that the first through hole 21 is closed by the first baffle plate 3, and the second through hole 22 is closed by the second baffle plate 4, as shown in the state of fig. 1; the design of this example can reduce the amount of culture medium and antibody used. When the antibody elution is performed, the first baffle 3 and the second baffle 4 are slid upwards to open the first through hole 21 and the second through hole 22, as shown in fig. 2, more liquid can be used for elution, which facilitates better elution of the antibody. After the elution is finished, the piston 5 is pulled out to pour out the liquid, and then the piston 5 is inserted into the culture dish through hole 11.

The device for the immunochemical fluorescent staining of the mesenchymal stem cell surface marker not only can reduce the using amount of a culture medium and an antibody, but also is simple and convenient to use, and greatly improves the quality and the efficiency of the immunochemical fluorescent staining of the mesenchymal stem cell.

The foregoing is a more detailed description of the present invention that is presented in conjunction with specific embodiments, and the practice of the invention is not to be considered limited to those descriptions. It will be apparent to those skilled in the art that a number of simple derivations or substitutions can be made without departing from the inventive concept.

Claims

1. A device for the immunochemical fluorescent staining of mesenchymal stem cell surface markers, which is characterized in that: comprises a culture dish main body (1) and a culture tank side wall (2);

the culture tank side wall (2) is arranged in the accommodating cavity of the culture dish main body (1), the culture tank side wall (2) is enclosed into an independent culture tank (20) in the accommodating cavity of the culture dish main body (1), and the culture tank side wall (2) is in a horn-shaped design with an upward opening;

the side wall (2) of the culture tank is provided with a first through hole (21) and a second through hole (22) which can be opened and closed at the symmetrical positions of the bottom of the side wall;

the bottom of the side wall of the culture dish main body (1) is provided with a culture dish through hole (11) capable of being opened and closed.

2. The apparatus of claim 1, wherein: the device also comprises a first baffle (3) and a second baffle (4);

the first baffle (3) is mounted on the side wall (2) of the culture tank in a vertically sliding manner and is mounted at the position of the first through hole (21); the first through hole (21) is opened or closed by moving the first baffle (3) up and down;

the second baffle (4) is arranged on the side wall (2) of the culture tank in a vertically sliding manner and is arranged at the position of the second through hole (22); the second through hole (22) is opened or closed by moving the second shutter (4) up and down.

3. The apparatus of claim 1, wherein: still include piston (5), piston (5) install in culture dish through-hole (11), realize closing or opening of culture dish through-hole (11) through the packing in or pulling out of piston (5).

4. The apparatus according to any one of claims 1-3, wherein: the culture tank (20) is located at the center of the accommodating cavity of the culture dish main body (1), and the center of the culture tank (20) is overlapped with the center of the accommodating cavity of the culture dish main body (1).

5. The apparatus according to any one of claims 1-3, wherein: the chamber that holds of culture dish main part (1) is cylindrically, culture tank (20) are enclosed by culture tank lateral wall (2) and close and are inverted cone frustum shape, and the axis that holds the chamber of culture dish main part (1) coincides with the axis of culture tank (20) of cone frustum shape.

6. The apparatus according to any one of claims 1-3, wherein: the culture dish main body (1) and the side wall (2) of the culture tank are both transparent.