Culture dish for laser confocal microscope

Abstract

The utility model relates to a culture dish, in particular to a culture dish for a laser confocal microscope, which comprises a dish body and a cover body, the dish body is a cylinder with a shooting groove in the top surface; the cover body comprises an outer cylinder and an inner cylinder; the inner column body is arranged in the outer column body, and the top surface of the inner column body is not lower than the top surface of the outer column body; the outer column body is provided with an annular groove surrounding the inner column body, and the inner column body is provided with a culture groove; the cover body is detachably connected with the dish body; and when the cover body covers the opening in the top surface of the dish body, the culture groove and the shooting groove are oppositely arranged. Compared with the prior art, the culture dish for the laser confocal microscope solves the problems that a conventional culture dish in the prior art does not meet the requirements of the laser confocal microscope, and an existing culture dish for the laser confocal microscope is complex to use and tedious to operate. According to the scheme, the culture dish for the laser confocal microscope, which can be repeatedly used, has relatively small limitation on samples and can be directly used for experiments such as invasion and the like, is realized.

Description

Technical Field

[0001] This utility model relates to a culture dish, specifically a culture dish for use in laser confocal microscopy. Background Technology

[0002] Laser Scanning Confocal Microscopy (LSCM) is a high-resolution optical imaging technique based on the principle of conjugate focusing. It enables three-dimensional, non-destructive observation of biological samples or materials through a laser light source, dual pinholes (illumination pinhole and probe pinhole), and a computer image processing system. Therefore, LSCM is widely used in biomedicine (e.g., dynamic observation of live cells, subcellular structure localization, tumor mechanism and drug screening research) and materials science (e.g., analysis of material surface morphology, three-dimensional quantitative analysis of metal corrosion / wear, microstructure characterization of MEMS devices).

[0003] Laser confocal microscopy can not only capture immunofluorescence images but also images of tissues with a certain thickness (especially PDO), forming a relatively complete and three-dimensional image record. This provides a holistic and objective three-dimensional representation of the relationships between different tissues, making it widely used in medicine and various other fields. However, each image capture using laser confocal microscopy requires the use of disposable imaging consumables specifically designed for object placement. These consumables have certain thickness requirements, resulting in high costs and numerous limitations on sample selection.

[0004] For example, CN101876748A discloses a reusable cell culture dish for laser confocal microscopy. The dish has a concave bottom with a light-transmitting hole, a lower pad, a coverslip, an upper pad, and a cell culture core. The cell culture core is sealed to the concave bottom of the cell culture dish. The upper pad, lower pad, and cell culture core all have light-transmitting holes of the same size and corresponding to the light-transmitting holes on the bottom of the cell culture dish. A cell culture dish lid covers the cell culture core. However, this design results in a complex structure and cumbersome operation.

[0005] For example, CN210103978U discloses a cell culture dish for easy labeling in laser confocal microscopy, and CN214174130U discloses a cell culture dish for laser confocal microscopy. Both require the use of glass slides / coverslips, which is cumbersome in experimental operation. The coverslips need to be cleaned and dried and sterilized before they can be used.

[0006] Therefore, there is a need for a culture dish suitable for laser confocal microscopy to meet the urgent need for reusability and ease of use. Utility Model Content

[0007] The purpose of this invention is to provide a culture dish for laser confocal microscopy to solve at least one of the aforementioned problems. This addresses the issues of conventional culture dishes in the prior art failing to meet the requirements of laser confocal microscopy, and the complexity and cumbersome operation of existing culture dishes for laser confocal microscopy. This solution provides a reusable culture dish for laser confocal microscopy that has relatively fewer restrictions on samples and allows for direct invasive experiments. During use, samples do not need to be transferred back and forth, thus avoiding sample damage and loss.

[0008] The objective of this utility model is achieved through the following technical solution:

[0009] A culture dish for use in laser confocal microscopy, comprising a dish body and a lid;

[0010] The dish body is a column with a shooting groove on its top surface;

[0011] The cover includes an outer column and an inner column; the inner column is disposed inside the outer column, and the top surface of the inner column is not lower than the top surface of the outer column; the outer column has an annular groove surrounding the inner column, and the inner column has a culture groove.

[0012] The lid and the dish body are detachably connected;

[0013] When the cover is placed over the opening on the top surface of the dish, the culture groove and the photography groove are positioned opposite each other.

[0014] Preferably, the column is a cylindrical structure, the outer column is a cylindrical structure, and the inner column is a cylindrical structure.

[0015] Preferably, the shooting groove is located at the center of the dish body, and the center line of the shooting groove coincides with the center line of the column body to form a concentric structure.

[0016] Preferably, the top surface of the inner column is higher than the top surface of the outer column.

[0017] Preferably, the bottom surface of the annular groove is lower than the bottom surface of the culture groove.

[0018] Preferably, the culture groove is located at the center of the inner column, and the center line of the culture groove coincides with the center line of the inner column to form a concentric structure; the inner column is located at the center of the outer column, and the center line of the inner column coincides with the center line of the outer column to form a concentric structure.

[0019] Preferably, the outer diameter of the inner column is not greater than the inner diameter of the column, and the outer diameter of the column is not greater than the inner diameter of the outer column;

[0020] When the lid is placed over the opening on the top surface of the dish, the side wall of the dish extends into the annular groove, thus sealing the culture groove and the imaging groove.

[0021] Preferably, it includes one or more of the following:

[0022] i) The height of the culture groove is 0.5-1mm;

[0023] ii) The inner diameter of the culture groove is 1.5-2 cm;

[0024] iii) The inner diameter of the annular groove is 2.5-4 cm.

[0025] Preferably, it includes one or both of the following:

[0026] i) The height of the culture groove is 0.8 mm;

[0027] ii) The inner diameter of the culture groove is 1.8 cm.

[0028] Preferably, it includes one or both of the following:

[0029] i) The material of the vessel is borosilicate glass;

[0030] ii) The cover is made of borosilicate glass.

[0031] The bottom surface of the dish is made of borosilicate glass, which can be used not only for laser confocal microscopy measurements, but also for sterilization by high temperature and high pressure.

[0032] The working principle of this utility model is as follows:

[0033] In use, the tissue / cells / organoids to be photographed are first cultured in the culture groove of the cover. After the culture is completed, the cover is closed on the opening of the dish, so that the cultured tissue / cells / organoids to be photographed fall into the imaging groove of the dish. Then, the tissue / cells / organoids to be photographed are fixed and stained inside the dish. Finally, laser confocal imaging can be performed directly.

[0034] Compared with the prior art, the present invention has the following beneficial effects:

[0035] The culture dishes used in this method eliminate the need for repeated sample transfers, effectively reducing sample loss and thus improving image quality, work efficiency, and simplifying the overall testing process.

[0036] The culture dishes prepared in this method can be directly autoclaved after use and can be reused. The processing operation is simple and easy.

[0037] The culture dish of this scheme has the advantages of simple structure, reasonable setup and low production cost. Attached Figure Description

[0038] Figure 1 A perspective view of the structure of a petri dish;

[0039] Figure 2 A perspective view of the structure of the petri dish;

[0040] Figure 3 A perspective view of the structure of the lid in a petri dish;

[0041] Figure 4 A perspective view of the structure of the petri dish with the lid inverted (during cultivation);

[0042] In the diagram: 1-vessel; 11-cylinder; 12-photographing groove; 2-lid; 21-outer cylinder; 22-inner cylinder; 23-annular groove; 24-culture groove. Detailed Implementation

[0043] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments.

[0044] Example 1

[0045] A culture dish for laser confocal microscopy, such as Figure 1-4 As shown, it includes a dish body 1 and a lid body 2;

[0046] The dish body 1 is a column 11 with a shooting groove 12 on its top surface;

[0047] The cover 2 includes an outer column 21 and an inner column 22; the inner column 22 is disposed inside the outer column 21, and the top surface of the inner column 22 is not lower than the top surface of the outer column 21; the outer column 21 has an annular groove 23 surrounding the inner column 22, and the inner column 22 has a cultivation groove 24.

[0048] The cover 2 and the dish 1 are detachably connected;

[0049] When the cover 2 is placed over the opening on the top surface of the dish 1, the culture groove 24 is positioned opposite to the imaging groove 12.

[0050] More specifically, in this embodiment:

[0051] This culture dish for laser confocal microscopy consists of two parts: the dish body 1 and the cover body 2. Figure 1 As shown, both adopt a cylindrical structure 11, wherein the height of the dish 1 is greater than the height of the lid 2, and the lid 2 can cover the top surface of the dish 1.

[0052] like Figure 2 As shown, the dish 1 is a cylinder 11 with a non-penetrating imaging groove 12 on its top surface. This imaging groove 12 is used to accommodate and place the tissue / cells / organoids to be imaged during laser confocal imaging. The imaging groove 12 is located at the center of the cylinder 11, making it concentric with the cylinder 11 (the centerline of the imaging groove 12 coincides with the centerline of the cylinder 11). The dimensions of the dish 1 are matched to the laser confocal microscope. To obtain better imaging results, the bottom thickness of the dish 1 can be controlled between 0.16-0.19 mm, and the wall thickness can be controlled between 0.5-1 mm.

[0053] like Figure 3 As shown, the cover 2 includes an outer column 21 and an inner column 22. The inner column 22 is disposed inside the outer column 21, and its top surface is higher than the top surface of the outer column 21. A culture groove 24, which does not penetrate the inner column 22, is formed on the top surface of the inner column 22. Simultaneously, an annular groove 23, surrounding the inner column 22, is formed on the top surface of the outer column 21 for matching the dish body 1. The culture groove 24 is used to culture the tissue / cells / organoids to be photographed before imaging to form collagen blocks. In this embodiment, the culture groove 24 is located at the center of the inner column 22, and the inner column 22 is located at the center of the outer column 21, such that the culture groove 24, the inner column 22, the annular groove 23, and the outer column 21 are all concentrically arranged (the center lines of the culture groove 24, the inner column 22, and the annular groove 23 all coincide with the center line of the outer column 21). Furthermore, in this embodiment, the bottom surface of the annular groove 23 is lower than the bottom surface of the culture groove 24, and the top surface of the inner column 22 is higher than the top surface of the outer column 21. This allows the culture groove 24 to have a larger culture space and minimizes the impact and contamination on the annular groove 23 during the operation of the culture groove 24.

[0054] To ensure that the lid 2 fits snugly over the opening on the top surface of the dish 1, as follows: Figure 1 As shown, the outer diameter of the inner column 22 should not be greater than the inner diameter of the column 11, and the outer diameter of the column 11 should not be greater than the inner diameter of the outer column 21. Thus, when the cover 2 is closed at the opening on the top surface of the dish 1, the top part of the side wall of the dish 1 extends into the annular groove 23 to facilitate the closing and removal of the cover 2, thereby sealing the internal space (culture groove 24 and imaging groove 12).

[0055] Based on the aforementioned structural limitations, in this embodiment, the height of the culture groove 24 is controlled within 0.5-1 mm, and its inner diameter is set within 1.5-2 cm. Simultaneously, the inner diameter of the annular groove 23 is controlled within the range of 2.5-4 cm. More preferably, the height of the culture groove 24 is set to 0.8 mm, and its inner diameter is set to 1.8 cm. At this size, it can be well matched with a laser confocal microscope, resulting in an optimal imaging range. Furthermore, in this embodiment, the dish body 1 can be made of conventional borosilicate glass, and the cover 2 is also made of borosilicate glass.

[0056] When using this petri dish:

[0057] (1) Select cover body 2 with different depths of culture groove 24 according to different tissues, cells or organoids (the dish body 1 can be used as a standard part).

[0058] (2) Taking cells or organoids as an example, first invert the operculum 2, such as... Figure 4 As shown, place the materials to be used for experiments and photography in culture groove 24 (resuspend cells or organoids in type I collagen and seed them into culture groove 24, adding enough to be level with the edge of culture groove 24). After the collagen solidifies, add culture medium and continue culturing in an incubator at 37°C (set the cell culture time according to the experimental purpose). After the cell culture is completed, gently aspirate the waste liquid, invert the cover 2 and place it on the top surface of the dish 1, so that culture groove 24 is opposite to photography groove 12. The collagen block obtained from the culture will slowly descend under the action of gravity and separate from the cover 2 (culture groove 24) (demolding). After the collagen block descends to the bottom of photography groove 12, perform pre-photography treatment steps such as fixation and staining on the entire collagen block. After the treatment is completed, laser confocal imaging can be performed directly.

[0059] (3) After use, wash the petri dish and autoclave it. It can then be used again.

[0060] Because the culture dish can select a suitable size cover 2 for culturing according to the type and size of the tissue / cell / organoid to be photographed, it can achieve the best imaging effect, thereby improving the success rate and reducing unnecessary repetitions.

[0061] The above description of the embodiments is provided to enable those skilled in the art to understand and use the utility model. It will be apparent to those skilled in the art that various modifications can be easily made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present utility model is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present utility model without departing from its scope should be within the protection scope of the present utility model.

Claims

1. A petri dish for laser confocal microscopy, characterized in that, It includes a dish body (1) and a lid body (2); The dish body (1) is a column (11) with a shooting groove (12) on its top surface; The cover (2) includes an outer column (21) and an inner column (22); the inner column (22) is disposed inside the outer column (21), and the top surface of the inner column (22) is not lower than the top surface of the outer column (21); the outer column (21) has an annular groove (23) surrounding the inner column (22), and the inner column (22) has a cultivation groove (24); The cover (2) and the dish (1) are detachably connected; When the cover (2) is placed over the opening on the top surface of the dish (1), the culture groove (24) is positioned opposite to the photographing groove (12).

2. The dish for laser confocal microscopy according to claim 1, characterized in that, The column (11) is a cylindrical structure, the outer column (21) is a cylindrical structure, and the inner column (22) is a cylindrical structure.

3. The dish for laser confocal microscopy according to claim 1, wherein The shooting groove (12) is located at the center of the dish body (1), and the center line of the shooting groove (12) coincides with the center line of the column body (11) to form a concentric structure.

4. The dish for laser confocal microscopy according to claim 1, wherein The top surface of the inner column (22) is set higher than the top surface of the outer column (21).

5. The dish for laser confocal microscopy according to claim 1, wherein The bottom surface of the annular groove (23) is set lower than the bottom surface of the culture groove (24).

6. A culture dish for laser confocal microscopy according to claim 1, characterized in that, The culture groove (24) is located at the center of the inner column (22), and the center line of the culture groove (24) coincides with the center line of the inner column (22) to form a concentric structure; the inner column (22) is located at the center of the outer column (21), and the center line of the inner column (22) coincides with the center line of the outer column (21) to form a concentric structure.

7. The dish for laser confocal microscopy according to claim 1, wherein The outer diameter of the inner column (22) is not greater than the inner diameter of the column (11), and the outer diameter of the column (11) is not greater than the inner diameter of the outer column (21); When the cover (2) is placed over the opening on the top surface of the dish (1), the side wall of the dish (1) extends into the annular groove (23), thereby sealing the culture groove (24) and the photographing groove (12).

8. The dish for laser confocal microscopy according to claim 1, wherein Includes one or more of the following: i) The height of the culture groove (24) is 0.5-1mm; ii) The inner diameter of the culture groove (24) is 1.5-2 cm; iii) The inner diameter of the annular groove (23) is 2.5-4 cm.

9. The dish for laser confocal microscopy according to claim 8, characterized in that Includes one or both of the following: i) The height of the culture groove (24) is 0.8 mm; ii) The inner diameter of the culture groove (24) is 1.8 cm.

10. The petri dish for laser confocal microscopy of claim 1, wherein, Includes one or both of the following: i) The material of the dish body (1) is borosilicate glass; ii) The cover (2) is made of borosilicate glass.

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