CN110791416A - Culture dish and culture dish manufacturing method - Google Patents

Abstract

The invention provides a culture dish and a culture dish manufacturing method, and belongs to the technical field of biochemistry. It has solved the not enough problem of adhesion when current culture dish cultivates the cell. The culture dish comprises a dish body and a climbing sheet, wherein the bottom wall of the dish body is provided with a through hole, the climbing sheet is fixed on the bottom surface of the dish body, and the climbing sheet covers the through hole; the climbing film is made of APET or cycloolefin copolymer (COC) materials and is subjected to plasma treatment. The culture dish manufacturing method comprises the following sequential steps, wherein in the first step, the materials are prepared, and the dish body, the climbing sheet and the dish cover are manufactured in no sequence; secondly, gluing; thirdly, assembling; fourthly, plasma treatment, namely putting the dish body and the slide assembly part into plasma treatment equipment for plasma treatment; and fifthly, post-processing. This culture dish is through the material of replacement slide, and attached and extension characteristic when both having improved cell culture guarantees optical characteristic and low dielectric constant again, improves cell imaging quality.

Description

Culture dish and culture dish manufacturing method

Technical Field

The invention belongs to the technical field of biochemistry, relates to a biochemical device, and particularly relates to a culture dish.

The invention belongs to the technical field of biochemistry, relates to a biochemical device, and particularly relates to a culture dish manufacturing method.

Background

Cell culture is the most central and basic technology in biotechnology; a petri dish is a laboratory vessel used for microbial or cell culture. The culture dish is usually composed of a flat disc-shaped bottom and a lid.

The glass-based culture dish is a cell culture dish with special purposes, and the limitation of materials at the bottom of a common culture dish cannot meet the special purposes of living cell imaging, laser confocal scanning imaging and the like. It is currently common to use a cover glass made of borosilicate glass for supporting microorganisms or cells in the middle area of the bottom of the culture dish.

Cell attachment and expansion are one of the important indicators in cell culture, and although attachment and expansion are the basic growth characteristics of most in vitro cultured cells, individual cells grow with poor attachment and expansion characteristics, such as lymphocytes. In order to improve the adhesion properties, special cell adhesion promoting substances such as laminin, fibronectin, type III collagen, serum expansion factor, etc. are used. The high borosilicate glass is subjected to vacuum plasma treatment under different conditions twice, so that the cell adsorption capacity is improved to a certain extent, but the actual efficiency is not ideal, in other words, special cells cannot grow normally on the glass-based culture dish, particularly, the adhesion and extension cannot be guaranteed, and further, the problem that the special cells cannot be subjected to related experiments is solved.

In addition, the existing glass culture dish has the defects that a great amount of chemical substances are needed in the production process of glass, and some chemical substances which can influence the growth of microorganisms or cells are inevitably remained on the surface of the climbing sheet.

The required culture conditions are not completely the same due to different characteristics of microorganisms or cells; part of microorganisms or cells need to be prevented from being polluted by mixed bacteria, so a microorganism culture dish (with an authorization notice number of CN 208472083U) is provided, and a sealing ring is arranged between a dish body and an upper cover to ensure that the culture dish is tightly combined. Some microorganisms or cells need good air permeability, so that a disposable culture dish (authorized bulletin No. CN 208218853U) is provided, wherein a plurality of rows of through holes are respectively arranged on the side walls of the dish body and the dish cover, and filtering membranes are arranged in the through holes. The existing culture dish has the defects of complex structure, high manufacturing cost, single function and the like.

Disclosure of Invention

The invention provides a culture dish, and aims to solve the technical problems of improving the attaching and stretching characteristics of the culture dish during cell culture and ensuring the subsequent treatment effects of microscope observation and the like.

The invention provides a culture dish, and aims to solve the technical problems of improving the attaching and stretching characteristics of the culture dish during cell culture and ensuring the subsequent treatment efficiency of microscope observation and the like.

The technical problem to be solved by the invention can be realized by the following technical scheme: a culture dish comprises a dish body and a climbing sheet, wherein the bottom wall of the dish body is provided with a through hole, the climbing sheet is fixed on the bottom surface of the dish body, and the climbing sheet covers the through hole; the climbing film is made of APET or cycloolefin copolymer (COC) materials and is subjected to plasma treatment.

The inventors only select APET or Cyclic Olefin Copolymer (COC) materials from a wide variety of resins, and the APET or Cyclic Olefin Copolymer (COC) materials may not contain chemical substances which influence cell growth, thereby improving the growth quality of cells in a culture dish. APET or Cyclic Olefin Copolymer (COC) materials have excellent optical performance and low dielectric constant; in summary, the optical properties such as light transmittance and refractive index of the material are nearly consistent compared with those of glass, so that the culture dish can be subjected to subsequent processing such as a fluorescence microscope, a phase contrast microscope, a confocal microscope, a differential interference contrast microscope, living cell imaging and Fluorescence In Situ Hybridization (FISH) and the subsequent processing effect is ensured.

After the materials adopted by the climbing film are subjected to plasma treatment, the hydrophilicity of the climbing film is obviously improved, the adherent growth probability of cells is further improved, the cells are tightly attached to the surface of the climbing film, and therefore the imaging quality of the cells during subsequent treatment is improved.

In the culture dish, the culture dish also comprises a dish cover, a concave cavity is arranged on the bottom surface of the dish cover, a plurality of bulges are arranged on the top surface of the concave cavity, and grooves which are in one-to-one correspondence with the bulges are arranged on the top surface of the dish body; when the dish cover is buckled on the dish body and the bulge is abutted against the top surface of the dish body, a first gap is formed between the top surface of the concave cavity of the dish cover and the top surface of the dish body; thereby ensuring the air permeability of the culture dish. When the dish cover is buckled on the dish body and the bulge is embedded into the groove, the top surface of the concave cavity of the dish cover and the top surface of the dish body depend on each other, and the air permeability between the inner cavity of the dish body and the outside is extremely poor.

The preparation method of the culture dish comprises the following sequential steps of preparing materials, manufacturing a dish body, a climbing sheet and a dish cover in no sequence, wherein the dish body and the dish cover are formed by resin injection molding, and cutting a sheet made of APET or Cyclic Olefin Copolymer (COC) materials into the climbing sheet; secondly, gluing, namely coating glue on the bottom surface of the dish body; thirdly, assembling, namely covering the climbing sheet at the through hole, and bonding the climbing sheet and the dish body by adopting glue, thereby forming a dish body climbing sheet assembly part; fourthly, plasma treatment, namely putting the dish body and the slide assembly part into plasma treatment equipment for plasma treatment; and fifthly, post-treatment, namely buckling the dish cover on the dish body, and carrying out packaging and disinfection treatment.

Compared with the prior art, the culture dish has the advantages that the material of the climbing sheet is replaced, so that the attaching and stretching characteristics during cell culture are improved, the optical characteristics and the low dielectric constant are ensured, and the cell imaging quality is improved.

The climbing sheet is formed by cutting a sheet made of APET or Cyclic Olefin Copolymer (COC) materials, and one sheet can be cut into a plurality of climbing sheets, so that the consistency of the climbing sheet quality, especially the consistency of the optical performance, is effectively ensured. The manufacturing method of the climbing sheet is suitable for automatic processing, not only ensures the cutting efficiency and the cutting quality, but also avoids the climbing sheet from being polluted, and further ensures that the culture dish is suitable for automatic production.

The culture dish is provided with the groove on the dish body and the bulge on the dish cover, so that the culture dish is suitable for culturing microorganisms or cells under different conditions; the groove is synchronously formed when the dish body is injection molded, and the bulge is synchronously formed when the dish cover is injection molded. The culture dish has the advantages of simple structure, high universality, wide application range and low manufacturing cost.

Drawings

FIG. 1 is a front view schematically showing the structure of a culture dish.

Fig. 2 and 3 are schematic views of the structure in different states along a-a in fig. 1.

Fig. 4 is a schematic perspective view of the culture dish in an open state.

Fig. 5 is a schematic sectional view of the dish body.

Fig. 6 is a schematic view of a structure for coating glue on the dish body.

Figure 7 is a schematic cross-sectional view of a capsule mounting assembly.

FIG. 8 shows fluorescence spotting effect of mCherry-GFP-LC3 protein expressed in cells photographed by glass-bottom culture dish laser confocal microscope.

FIG. 9 shows fluorescence spotting effect of the slide by using a culture dish laser confocal microscope made of APET material to shoot mCherry-GFP-LC3 protein expressed in cells.

In the figure, 1 is a dish body; 1a, a groove; 1b, a through hole; 1c, positioning a groove; 2. a dish cover; 2a, a concave cavity; 2b, a bulge; 3. climbing sheets; 4. a first gap; 5. a second gap; 6. and (4) glue.

Detailed Description

The following are specific embodiments of the present invention and are further described with reference to the drawings, but the present invention is not limited to these embodiments.

As shown in fig. 1 to 4, a culture dish includes adish body 1, a dish lid 2, and aslide 3.

The bottom wall of thedish body 1 is provided with athrough hole 1b, the bottom surface of thedish body 1 is provided with apositioning groove 1c, and the outer side surface of the top of thedish body 1 is conical.

The dish cover 2 can be buckled on the top of thedish body 1, aconcave cavity 2a is formed in the bottom surface of the dish cover 2, a plurality ofbulges 2b are formed in the top surface of theconcave cavity 2a, the number of thebulges 2b is three according to the drawing in the specification, and the number of thebulges 2b can be adaptively increased or decreased according to actual conditions. Thebulges 2b are uniformly distributed along the circumferential direction of the axial lead of the dish cover 2, and the height of thebulges 2b is less than 2 mm. The top surface of thedish body 1 is provided with grooves 1a which are in one-to-one correspondence with thebulges 2b, the width of each groove 1a is matched with that of thebulge 2b, and the depth of each groove 1a is matched with that of thebulge 2 b. The side surface of theconcave cavity 2a of the dish cover 2 is also conical, and the diameter of the outer side surface at the top of thedish body 1 is matched with the diameter of the side surface of theconcave cavity 2a of the dish cover 2, so that the dish cover 2 is more easily buckled on thedish body 1.

As shown in fig. 2, when the dish cover 2 is buckled on thedish body 1 and theprotrusion 2b is embedded into the groove 1a, the top surface of thecavity 2a of the dish cover 2 is leaned against the top surface of thedish body 1, the outer side surface of the top of thedish body 1 is basically inosculated and jointed with the side surface of thecavity 2a of the dish cover 2, and the air permeability between the inner cavity of thedish body 1 and the outside is extremely poor.

As shown in fig. 3, when the dish cover 2 is buckled on thedish body 1 and thebulge 2b is abutted against the top surface of thedish body 1, a first gap 4 is formed between the top surface of thecavity 2a of the dish cover 2 and the top surface of thedish body 1, and a second gap 5 which enables the first gap 4 to be communicated with the outside is also formed between the outer side surface of the top of thedish body 1 and the side surface of thecavity 2a of the dish cover 2, so that the air permeability of the culture dish is ensured.

Theclimbing sheet 3 is in a circular sheet shape, the thickness of theclimbing sheet 3 is 0.16 mm-0.19 mm, and theclimbing sheet 3 is made of APET or cycloolefin copolymer COC material. Theclimbing piece 3 is positioned in thepositioning groove 1c of thedish body 1, and theclimbing piece 3 and thedish body 1 are bonded and fixed by glue 6.

The advantages of the culture dish are further illustrated by illustrating the above method of manufacturing the culture dish as shown in fig. 1 to 7. Preparing materials; thedish body 1, theclimbing sheet 3 and the dish cover 2 are manufactured in no sequence. Thedish body 1 and the dish cover 2 are both formed by injection molding of polystyrene resin, so that the automatic production can be realized, the production efficiency is high, and the pollution to thedish body 1 and the dish cover 2 can be avoided. The recess 1a is formed when thecapsule 1 is injection moulded and theprotrusion 2b is formed when the capsule 2 is injection moulded. Theclimbing piece 3 is formed by cutting a sheet made of APET or cycloolefin copolymer COC material, and the diameter of theclimbing piece 3 is matched with that of thepositioning groove 1 c.

And secondly, gluing, namely coating glue 6 on the bottom surface of thepositioning groove 1c of thedish body 1, as shown in figure 6.

Thirdly, assembling, namely placing theclimbing piece 3 in thepositioning groove 1c, so that theclimbing piece 3 covers the throughhole 1b, and bonding theclimbing piece 3 and thedish body 1 by using the glue 6 after the glue 6 is solidified; thecapsule 1 is formed as aclimbing sheet 3 assembly, as shown in figure 7.

The fourth step of plasma treatment, namely putting the assembly part of theclimbing piece 3 of thedish body 1 into plasma treatment equipment for plasma treatment; in other words, the surface hydrophilicity of theslide 3 is obviously increased after the plasma treatment, so that theslide 3 is more suitable for the adherent growth of cells.

And fifthly, post-treatment, namely buckling the dish cover 2 on thedish body 1 and carrying out packaging and disinfection treatment.

As shown in fig. 8 and 9, the optical properties of the material of theslide 3 in the culture dish are nearly the same as those of glass, compared with the conventional glass-based culture dish or the glass-based culture dish subjected to plasma treatment, but the hydrophilicity of theslide 3 in the culture dish produced by the above-mentioned manufacturing method is significantly improved, and particularly special cells such as lymphocytes can be attached to the surface of theslide 3 to grow, in other words, when the special cells are cultured by the culture dish produced by the above-mentioned manufacturing method, the cells are attached to the surface of theslide 3 and the cells are fully stretched. Therefore, the cell imaging quality is higher during the post-treatment of a fluorescence microscope, a phase contrast microscope, a confocal microscope, a differential interference contrast microscope, living cell imaging, Fluorescence In Situ Hybridization (FISH) and the like; and further, the problem that the special cells cannot normally grow on a glass medium culture dish to cause that the special cells cannot carry out related experiments is avoided.

Claims (10)

1. A culture dish comprises a dish body (1) and a climbing sheet (3), wherein a through hole (1 b) is formed in the bottom wall of the dish body (1), the climbing sheet (3) is fixed on the bottom surface of the dish body (1), and the climbing sheet (3) covers the through hole (1 b); the climbing film is characterized in that the climbing film (3) is made of APET or cycloolefin copolymer (COC) material, and the climbing film (3) is subjected to plasma treatment.

2. The culture dish according to claim 1, further comprising a dish cover (2), wherein a concave cavity (2 a) is formed in the bottom surface of the dish cover (2), a plurality of bulges (2 b) are formed on the top surface of the concave cavity (2 a), grooves (1 a) corresponding to the bulges (2 b) one by one are formed in the top surface of the dish body (1), and when the dish cover (2) is buckled on the dish body (1) and the bulges (2 b) are abutted against the top surface of the dish body (1), a gap (4) is formed between the top surface of the concave cavity (2 a) of the dish cover (2) and the top surface of the dish body (1); when the dish cover (2) is buckled on the dish body (1) and the bulge (2 b) is embedded into the groove (1 a), the top surface of the concave cavity (2 a) of the dish cover (2) is leaned against the top surface of the dish body (1).

3. Dish according to claim 2, characterized in that the width of the grooves (1 a) coincides with the width of the protrusions (2 b).

4. Dish according to claim 2, characterized in that the depth of the grooves (1 a) coincides with the height of the protrusions (2 b).

5. Culture dish according to claim 2 or 3 or 4, characterized in that the outer side of the top of the dish body (1) and/or the side of the cavity (2 a) of the dish lid (2) is tapered.

6. The culture dish according to claim 5, wherein the diameter of the outer side surface of the top of the dish body (1) is matched with the diameter of the side surface of the concave cavity (2 a) of the dish cover (2); when the dish cover (2) is buckled on the dish body (1) and the bulge (2 b) is propped against the top surface of the dish body (1), a second gap (5) which enables the first gap (4) to be communicated with the outside is arranged between the outer side surface of the top of the dish body (1) and the side surface of the concave cavity (2 a) of the dish cover (2).

7. The culture dish according to claim 6, wherein when the dish cover (2) is buckled on the dish body (1) and the protrusion (2 b) is embedded in the groove (1 a), the outer side surface of the top of the dish body (1) is attached to the side surface of the concave cavity (2 a) of the dish cover (2).

8. A culture dish according to claim 2 or 3 or 4, wherein the height of the protrusions (2 b) is less than 2 mm.

9. A method of manufacturing a culture dish according to any one of claims 1 to 8, wherein the method of manufacturing a culture dish is performed in the following sequence of steps,

firstly, preparing materials, namely manufacturing a dish body (1), a climbing piece (3) and a dish cover (2) in no sequence, wherein the dish body (1) and the dish cover (2) are formed by resin injection molding, and a sheet made of APET or Cyclic Olefin Copolymer (COC) materials is cut into the climbing piece (3);

secondly, gluing, namely coating glue (6) on the bottom surface of the dish body (1);

thirdly, assembling, namely covering the through hole (1 b) with the climbing sheet (3), and bonding the climbing sheet (3) and the dish body (1) by adopting glue (6), thereby forming an assembling part of the climbing sheet (3) of the dish body (1);

fourthly, plasma treatment, namely putting the dish body (1) climbing piece (3) assembly part into plasma treatment equipment for plasma treatment;

and fifthly, post-treatment, namely buckling the dish cover (2) on the dish body (1) and carrying out packaging and disinfection treatment.

10. Dish making method according to claim 9, characterised in that the thickness of the sheet is between 0.16mm and 0.19mm and the thickness of the climbing piece (3) is between 0.16mm and 0.19 mm.

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