CN218115467U

CN218115467U - Detachable cell culture box for microscope observation

Info

Publication number: CN218115467U
Application number: CN202222488701.7U
Authority: CN
Inventors: 李世武; 罗雄剑; 李明
Original assignee: Kunming Institute of Zoology of CAS
Current assignee: Kunming Institute of Zoology of CAS
Priority date: 2022-09-20
Filing date: 2022-09-20
Publication date: 2022-12-23
Anticipated expiration: 2032-09-20

Abstract

The utility model discloses a detachable cell culture box that supplies microscopic observation to use comprises box body and lid, the box body constitute by box bottom and box wall detachably, the box body be the cuboid, the minor face both sides of box bottom are equipped with the extension end, the peripheral integrated into one piece of box bottom be equipped with the seal groove, the seal groove in be equipped with the elastic sealing rubber circle, box wall bottom edge integrated into one piece have with the sealed groove suit sealed sealing strip of cooperation. The device solves the defects of poor adherent culture, cell aggregation, high price, inconvenience for fixation and complex operation of cell culture adherent culture of related products in the prior art, and has the advantages of simple manufacture, low cost, good adherent growth effect of cells, simple and convenient operation and wide market prospect.

Description

Detachable cell culture box for microscope observation

Technical Field

The utility model relates to a cell culture technical field protects a cell culture box, especially a detachable cell culture box that supplies microscopic observation to use.

Background

In recent years, with the rising of brain science, it is an indispensable research method to compare and observe the fine structure of nerve cells (such as dendritic branches of neurons, dendritic spine morphology, mitochondria, cell membranes, etc.) in the field of neural research, and related products such as: nunc-Lab-Tek II Chamber Slide system (cat # 154534 PK) of thermofisher, glass-bottom culture dishes (cat # 801002, 801001) and glass-bottom culture plates (cat # 801004, 801006) of Wuxi Kangsu Limited, however, the price of the related products is high, and glass materials are adopted as the cell growth wall surface. Because special treatment (Poly-D-lysine (PDL) and Laminin (Laminin) coating) is needed when nerve cells (such as nerve stem cells and neurons) are cultured in an adherent manner, the requirement on experimenters is higher in the experimental operation process. Due to the difference (unevenness) in the coating process, the glass shows that the smooth and hydrophobic surfaces cause poor adherent culture effect of nerve cells (neural stem cells, neurons and the like), the phenomenon of cell aggregation seriously influences the experimental result, and further causes the waste of scientific research funds. In addition, the microstructure of neuronal cell needs to be observed under microscope high power lens (60-100 times), because the distance between the microscope high power objective lens and the target object to be photographed is 0.15-0.19mm, the conventional technique is to use the Slide in the cell culture dish/plate or use the Nunc ™ Lab-Tek II Chamber Slide system and the glass bottom culture dish/plate mentioned above to perform plating culture after being coated, the former needs to be processed by immunofluorescence experiment to be photographed by the laser confocal instrument, while the latter can directly observe the cell on the laser confocal instrument with high power lens, but the price is expensive, which is not suitable for large scale experiment. When a researcher adopts a slide climbing strategy, in the experimental process, as foreign matters exist in the slide occasionally and the slide is easy to float in the culture solution, the slide is not fully contacted with the culture solution, and if more than two slides or slides are placed, the slides or the slides are easy to overlap, so that the cell culture is influenced; on the other hand, the slide for immunofluorescence experiments currently available on the market is usually circular, the diameter is 2.5cm at most, when the slide is laid in a 6-well culture plate, a large area of the bottom of the well is not used for the experiment, so that the slide is easy to slide and float in the well, and the cell culture is seriously influenced. In addition, since the efficiency of neuron transfection is very low, a large area of adherent neuron culture is required to increase the number of successful transfection, and a slide with a diameter of 2.5cm is relatively insufficient. It is worth mentioning that the schemes adopted by the prior common technologies all lead nerve cells (nerve stem cells, neurons and the like) to grow in an adherent manner on glass, which puts higher requirements on the manufacturing process and the coating method of the slide, and because the above mentioned disadvantages that the glass material is easy to gather for adherent culture of nerve cells and the like, the success rate of adherent culture of nerve cells (nerve stem cells, neurons and the like) is still low, and the waste of reagents and manpower is caused.

There is no prior art device for solving the above technical problems. Patent No. 202020728700.3 discloses a cell culture apparatus for a microscope, which is to prevent the problem that the temperature of the inside air fluctuates due to the fact that the concentration of carbon dioxide inside is too high and needs to be exchanged with the outside air, which leads to cell death or slows down the culture speed. Patent No. CN201820747709.1 discloses a cell culture apparatus for a microscope, which adopts a cell observation cover, is provided with a temperature controller to provide a constant temperature observation environment for cell culture, avoids mist generation, and is convenient to operate and observe. By adopting the cell gas control assembly, the gas concentration in the cell culture chamber can be detected, and the gas content required by cell culture can be reasonably controlled. Simultaneously the dish of culture dish is covered the design of recess, has reduced the air space of culture solution top inside the culture dish, can effectively prevent the water smoke formation of recess bottom to be convenient for observe. These devices solve different technical problems, adopt different technical schemes, and have different applicable conditions, cells and effects.

Therefore, a detachable cell culture box which is economical, easy and convenient to operate and available for microscopic observation is needed, and the cell culture box is suitable for adherent culture of nerve cells (neurons and neural stem cells) and convenient for immunofluorescence experiments and laser microscope photographing.

SUMMERY OF THE UTILITY MODEL

The utility model provides a detachable cell culture box convenient for microscopic observation to make up the defects of the prior art.

The utility model adopts the technical proposal that:

the utility model provides a supply detachable cell culture box of microscopic observation usefulness, comprises box body and lid, the box body constitute by box bottom and box wall detachably, the box body be the cuboid, the minor face both sides of box bottom are equipped with the extension end, the peripheral integrated into one piece of box bottom be equipped with the seal groove, the seal groove in be equipped with the elastic seal rubber circle, box wall bottom edge integrated into one piece have with the sealed groove sealing strip that suits the cooperation. The structure can ensure the normal work of the cell culture box, and is easy to disassemble and free from liquid leakage.

Further, the box bottom is made of polypropylene or other materials used for cell culture.

Preferably, the bottom of the box is made of a high transparent Tissue Culture (TC) treated polypropylene material for better cell adherent culture effect.

Preferably, the height of the box wall is 10-15mm.

Preferably, for the convenience of microscopic cell observation, the width of the box bottom is 20-25mm, the length of the box bottom is 40-50mm, and the thickness of the box bottom is 1-2mm.

Preferably, the size of the box cover is matched with that of the box body, the depth of the box cover is 10-15mm, and in order to take out the box cover, the middle parts of the two sides of the box cover are also provided with gripping parts which are convenient to grip.

In order to further strengthen the connection between the box bottom and the box wall and avoid liquid leakage, the four corners of the box wall are also provided with reversible buckles, and the box bottom is provided with buckle bosses 41 which are adaptive to the buckles.

Preferably, in order to facilitate the observation of microscopic cells and to prevent the relevant liquid reagents (such as fixing liquid, penetrating agent, sealing liquid, antibody liquid and sealing agent) from flowing away easily during immunofluorescence, the height of the two sides of the sealing agent in the sealing groove is not more than 1mm. Beyond 1mm, microscopic observation of the cells may be affected.

The working principle is as follows:

when the device works, a culture box is taken out firstly, then a box bottom is coated by using a high molecular reagent (PDL and/or Lamin) to enable the surface of the box bottom to become rough, whether the tightness between the box bottom and a box wall is good or not is detected according to whether a coating liquid seeps out, nerve cells are cultured on the surface of the box bottom in an adherent way after the tightness test is passed, after an experiment (such as immunofluorescence and the like) is finished, a buckle can be opened, the box wall is detached, a proper amount of sealing liquid is coated on the box bottom, a cover glass with a proper size and a proper thickness of 0.15mm is covered for sealing, finally, the extension end of the box bottom is fixed on a laser microscope fixing frame, and the corresponding fine structure of the nerve cells growing in the adherent way is observed.

Compared with the prior art, the utility model discloses following beneficial effect has:

1. the device overcomes the defects of poor adherent culture, cell aggregation and high price of the related product cell culture in the prior art, has simple manufacture, low cost, good adherent growth effect of the cells, simple and convenient operation and wide market prospect.

2. The device can be directly used for culturing nerves or other cells, and the polyethylene material processed by the highly transparent TC adopted at the bottom of the box has lower coating requirements and good cell adherent culture effect; the box bottom is used as a nerve cell wall-attaching surface, a cell-used slide is not required to be purchased, the problem that the slide floats or folds in a culture solution is solved, the area of the culture box is large, the number of cultured cells is large, the successful number of nerve cell transfection can be increased conveniently, and more cell numbers can be observed; the extending end of the box bottom can be fixed on a laser microscope fixing frame, and the micro structure related to the nerve cells growing adherent to the box bottom can be observed; the sealing groove is integrally formed at the periphery of the box bottom, and is beneficial to preventing related liquid reagents (such as fixing liquid, penetrating agent, sealing liquid, antibody liquid and sealing agent) from flowing away easily during immunofluorescence.

Drawings

FIG. 1 is a schematic view of the overall structure of the apparatus;

FIG. 2 is a schematic diagram of the device in a disassembled structure;

FIG. 3 is a schematic view of the overall structure of the device with a gripping member;

FIG. 4 is a schematic view of the present device with a gripping member in a disassembled configuration;

1-an extension end; 2-sealing groove; 3-elastic sealing rubber ring; 4-box bottom; 5-box walls; 6-box body; 7-box cover; 41-a snap boss; 51-buckling; 71-a grip.

Detailed Description

The present invention will be described in detail with reference to the accompanying drawings and specific embodiments, but the present invention is not limited thereto.

Example (b):

the utility model provides a supply detachable cell culture box of microscopic observation usefulness, comprises box body 6 and lid 7, box body 6 constitute by

box bottom

4 and 5 detachably of box wall, box body 6 be the cuboid, the minor face both sides of box bottom 4 are equipped with

extension end

1, 4 peripheral integrated into one piece of box bottom be equipped with seal groove 2, seal groove 2 in be equipped with elastic sealing rubber circle 3, box wall bottom edge integrated into one piece have with the sealed sealing strip of seal groove adaptation cooperation seal. The cell culture box can work normally, is easy to disassemble and does not leak liquid.

Further, the box bottom 4 is made of glass or a polypropylene material.

Preferably, the bottom 4 is made of a high transparent cell TC (Tissue culture, TC) treated polypropylene material for better cell attachment culture effect.

Preferably, the height of the box wall 5 is 10-15mm.

Preferably, for the convenience of microscopic cell observation, the width of the box bottom 4 is 20-25mm, the length of the box bottom 4 is 40-50mm, and the thickness of the box bottom 4 is 1-2mm.

Preferably, the size of the box cover 7 is matched with that of the box body 6, the depth of the box cover 7 is 10-15mm, and in order to facilitate taking out the box cover, the middle parts of two sides of the box cover 7 are also provided with gripping pieces 71 which are convenient to grasp.

In order to further strengthen the connection between the box bottom 4 and the box wall 5 and avoid liquid leakage, the four corners of the box wall 5 are also provided with reversible buckles 51, the buckles 51 are buckled on the box bottom 4 in a turnover way, and the box bottom is provided with buckle bosses 41 which are matched with the buckles.

The working principle is as follows: for adherent culture of nerve cells (e.g. neural stem cells, neurons, etc.):

when the device works, firstly, the culture box is taken out, the box bottom is coated by using a high molecular reagent (PDL and/or Lamin), the surface of the culture box becomes rough, whether the tightness of the box bottom and the box wall is good or not is detected according to whether the coating liquid seeps out, the nerve cells are cultured on the surface of the box bottom in an adherent way after the tightness test is passed, after the experiment (such as immunofluorescence and the like) is completed, the buckle can be opened, the box wall is detached, a proper amount of mounting liquid is coated on the box bottom, a cover glass with proper size and thickness of 0.15mm is covered for mounting, finally, the extension end of the box bottom is fixed on a laser microscope or electron microscope fixing frame, and the corresponding microstructure of the nerve cells growing in the adherent way is observed.

2. The device can be directly used for culturing nerves or other cells, and the polyethylene material processed by the highly transparent TC adopted at the bottom of the box has lower coating requirements and good cell adherent culture effect; the bottom of the culture box is used as the wall surface for attaching the nerve cells, so that the cell climbing sheet does not need to be purchased, the problem that the climbing sheet floats or folds in a culture solution is solved, the culture box has large area and more cultured cells, the successful number of transfection of the nerve cells is conveniently increased, and more cells can be observed; the extending end of the box bottom can be fixed on a laser microscope fixing frame, and the micro structure related to the nerve cells growing adherent to the box bottom can be observed; the sealing groove is integrally formed at the periphery of the box bottom, and is beneficial to preventing related liquid reagents (such as fixing liquid, penetrating agent, sealing liquid, antibody liquid and sealing agent) from flowing away easily during immunofluorescence.

Claims

1. The utility model provides a supply detachable cell culture box of microscope observation use, comprises box body (6) and lid (7), its characterized in that: box body (6) constitute by box bottom (4) and box wall (5) detachably, box body (6) be the cuboid, the minor face both sides of box bottom (4) are equipped with extension end (1), the peripheral integrated into one piece of box bottom be equipped with seal groove (2), seal groove (2) in be equipped with elastic sealing rubber circle (3), box wall (5) bottom edge integrated into one piece have with seal groove (2) the sealed sealing strip of cooperation that suits.

2. A removable cell culture cassette for microscopic observation according to claim 1, wherein: the box bottom (4) is made of a polypropylene ethylene material or other materials used for cell culture.

3. A removable cell culture cassette for microscopic observation according to claim 2, wherein: the box bottom (4) is made of a high-transparency tissue culture treated polypropylene material.

4. A removable cell culture cassette for microscopic observation according to claim 1, wherein: the height of the box wall (5) is 10-15mm.

5. A removable cell culture cassette for microscopic observation according to claim 1, wherein: the width of the box bottom (4) is 20-25mm, the length of the box bottom is 40-50mm, and the thickness of the box bottom is 1-2mm.

6. A removable cell culture cassette for microscopic observation according to claim 1, wherein: the size of the box cover (7) is matched with that of the box body (6), the depth of the box cover is 10-15mm, and the middle parts of the two sides of the box cover (7) are also provided with gripping parts (71) which are convenient to grip.

7. A removable cell culture cassette for microscopic observation according to claim 1, wherein: the four corners of the box wall (5) are also provided with reversible buckles (51), and the buckle turnably buckles buckle bosses (41) which are arranged on the box bottom (4) and are matched with the buckles.

8. A removable cell culture cassette for microscopic observation according to claim 1, wherein: the height of the sealing groove (2) is not more than 1mm.