WO2023040631A1

WO2023040631A1 - Novel double-sided solid culture dish and use thereof

Info

Publication number: WO2023040631A1
Application number: PCT/CN2022/115025
Authority: WO
Inventors: 田新朋; 石松标; 张敬; 龙丽娟
Original assignee: 中国科学院南海海洋研究所; 南方海洋科学与工程广东省实验室(广州)
Priority date: 2022-04-12
Filing date: 2022-08-26
Publication date: 2023-03-23
Also published as: CN218115429U

Abstract

Disclosed in the present application are a novel double-sided solid culture dish and use thereof, which relate to the fields of microculture, etc. The culture dish comprises an upper cover and a lower cover, a straight cylinder dish wall being arranged between the upper cover and the lower cover, and the upper cover, the lower cover and the straight cylinder dish wall forming a snap-fit structure; and a gauze type permeable membrane base, a grid-shaped fixing structure being arranged on the gauze type permeable membrane base, and "T"-shaped column structures being vertically arranged on joints or grid lines of the grid-shaped fixing structure. Culture dishes are formed between the gauze type permeable membrane base and the upper cover, and between the gauze type permeable membrane base and the lower cover respectively, and a solidified culture medium is made into the stable double-sided solid culture dish since the "T"-shaped column structures on the gauze type permeable membrane base and a circle of boss structure perpendicular to the wall of the straight cylinder dish wall jointly support a solid culture medium to be not separated from the gauze type permeable membrane base. According to the culture dish, double-sided preparation and the use of the solid culture medium and information exchange and interaction of double-sided microorganisms are achieved, such that related researches of non-contact interaction between microorganisms or interaction between the microorganisms and an environment become simple, convenient, rapid and efficient.

Description

A new type of double-sided solid culture dish and its application

Technical field:

The utility model relates to the preparation of double-sided solid culture medium in the fields of microbial cultivation, activity screening, biomedical detection, microbial ecology, and other related scientific research and teaching, in particular to a medium for easily and efficiently displaying the interaction between microorganisms on both sides Double-sided solid permeable Petri dishes.

Background technique:

Petri dishes are simple and important professional experimental supplies necessary for experiments, detection, and analysis related to research fields such as microbiology, cell biology, and medicine. The solid medium prepared by Petri dishes is the most basic and important operation in experiments related to microorganism and cell culture.

Referring to Figure 1, the traditional petri dish has only one groove, and a culture surface is prepared, and cultures such as inoculating microorganisms can only grow on the surface of a single side of the culture medium; severely limiting the ability of microorganisms on the culture medium surface to communicate with other types of microorganisms or Nutrient-matrix interaction studies.

Utility model content:

The purpose of this utility model is to design for the shortcomings of existing culture dishes such as low efficiency of selective separation and screening of functional microorganisms, cumbersome non-contact interaction experiments of microorganisms and easy pollution, and poor "simulation of in-situ environment" in microecological research. A new double-sided solid petri dish. The petri dish can realize the cultivation of microbial cells on both sides of the solid medium, can realize the efficient and selective separation and screening of functional microorganisms with antibacterial activity, and can prepare "simulated in situ environment" separation medium, etc., so that microorganisms can be non-contact Interaction, microbe-environment interaction and other related studies have become feasible and have the advantages of simplicity, efficiency, and low cost.

In order to achieve the above object, the utility model can take the following technical solutions:

A double-sided solid culture dish comprising:

cover;

lower lid;

A straight vessel wall, which is arranged between the upper cover and the lower cover, wherein a gauze-type transparent membrane bottom with a horizontal structure is arranged inside the straight vessel wall, and the upper and lower sides of the gauze-type transparent membrane bottom are Both can be used for cultivating microorganisms, and the upper cover and the lower cover are correspondingly fastened to the upper and lower end surfaces of the straight vessel wall.

As for the above-mentioned double-sided solid culture dish, further, the bottom of the gauze-type permeable membrane has a grid-like fixing structure, and the upper and lower sides of the grid-like fixing structure are respectively provided with several "T"-shaped column structures, and the "T"-shaped The column structure is perpendicular to the bottom of the gauze-type permeable membrane.

As for the above-mentioned double-sided solid culture dish, further, the "T"-shaped column structure includes: a column vertically connected to the bottom of the gauze-type permeable membrane, and a horizontal column vertically connected to the end of the column pole or plane.

As for the above-mentioned double-sided solid culture dish, further, the gauze-type permeable membrane bottom is supported by a permeable membrane and a grid-shaped fixed structure, and the "T"-shaped column structure is arranged at the junction of the grid-shaped fixed structure. dots or grid lines.

For the above-mentioned double-sided solid culture dish, further, a circle of protrusions is respectively provided above and below the contact position of the inner wall of the straight cylindrical dish wall close to the bottom of the gauze-type permeable membrane.

For the above-mentioned double-sided solid petri dish, further, the inner wall of the mouth edge of the upper cover and the lower cover is provided with a plurality of spherical protrusions, and a circle of spherical protrusions is provided outside the upper and lower mouth edges of the straight vessel wall. The spherical protruding point cooperates with the spherical protruding ring to form a buckle structure.

In the above-mentioned double-sided solid culture dish, further, the gauze-type permeable membrane bottom is arranged in the middle of the straight vessel wall, and the gauze-type permeable membrane bottom is integrated with the straight vessel wall.

For the double-sided solid culture dish as described above, further, the outer wall of the straight vessel is provided with a quantitative scale, the quantitative scale is perpendicular to the bottom of the gauze-type permeable membrane, and the zero scale of the quantitative scale is located at the quantitative The middle position of the ruler is aligned with the bottom of the gauze type permeable membrane.

In the above-mentioned double-sided solid culture dish, further, the inner diameter of the upper cover and the lower cover is larger than the outer diameter of the straight vessel wall, and the difference between the two diameters is kept within a set range.

As for the above-mentioned double-sided solid petri dish, further, there is a ventilation gap between the upper cover and the lower cover and the wall of the straight tube respectively.

Compared with the prior art, the utility model has the beneficial effects of:

1. The utility model changes the defect that the traditional culture dish can only prepare one-sided culture medium for microbial cell growth, and can satisfy the simultaneous growth of microorganisms on both sides of the solid culture medium, and can carry out material growth through the gauze-type permeable membrane bottom. and information exchange, etc.; the petri dish can realize non-contact interaction research between microorganisms or between microorganisms and other substrates, high-throughput selective separation and screening of microbial resources that produce active substances, etc., and the "in situ environment" of uncultivated microorganisms Important cutting-edge scientific research such as "simulation" training.

2. The utility model has the characteristics of simple and smart design, low cost, simple and fast, strong practicability, and various functions, which is beneficial to promote the research progress in the field of natural microbial resource mining.

Description of drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings that need to be used in the embodiments will be briefly introduced below. The drawings in the following description are only some embodiments of the application. Ordinary technicians can also obtain other drawings based on these drawings on the premise of not paying creative work.

Fig. 1 is the structural representation of prior art petri dish;

Fig. 2 is the cross-sectional structure schematic diagram of double-sided solid culture dish of the present utility model;

Fig. 3 is the top plan view schematic diagram of double-sided solid culture dish of the present invention;

Fig. 4 is a three-dimensional structural schematic diagram of a double-sided solid culture dish of the present invention in a closed state;

Fig. 5 is a three-dimensional structural schematic diagram of a double-sided solid culture dish of the present invention in a decomposed state;

Fig. 6 is a sectional view of a cut state of the double-sided solid culture dish of the present invention.

In the figure: 1. The lid of the culture dish; 2. The bottom groove of the culture dish; 3. The upper cover; 4. The lower cover; 5. The bottom of the gauze type permeable membrane; 1. Annular protruding platform structure; 9. Scale; 10. Spherical protruding point; 11. Spherical protruding ring; 12. Ventilation gap.

Detailed ways:

The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the application, not all of them. . Based on the embodiments in this application, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the scope of protection of this application.

Example

It should be noted that the terms "comprising" and "having" and any variations thereof in the embodiments of the present utility model are intended to cover non-exclusive inclusion, for example, processes, methods, systems, and products that include a series of steps or units Or the apparatus is not necessarily limited to those steps or units explicitly listed, but may include other steps or units not explicitly listed or inherent to the process, method, product or apparatus.

It is to be understood that the terms "center", "longitudinal", "transverse", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "Right", "Vertical", "Horizontal", "Top", "Bottom", "Inner", "Outer", "Clockwise", "Counterclockwise", "Axial", "Radial", " The orientation or positional relationship indicated by "circumferential direction" is based on the orientation or positional relationship shown in the drawings, and is only for the convenience of describing the utility model and simplifying the description, rather than indicating or implying that the referred device or element must have a specific orientation , are constructed and operated in a specific orientation and therefore cannot be construed as limiting the invention.

In the description of the present utility model, "plurality" means at least two, such as two, three, etc., unless otherwise specifically defined. In addition, unless otherwise clearly stipulated and limited, the terms "mounted", "connected" and "connected" should be interpreted in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical A connection can also be an electrical connection; it can be a direct connection or an indirect connection through an intermediary, and it can be the internal communication of two components. Those of ordinary skill in the art can understand the specific meanings of the above terms in the present invention in specific situations.

In the present invention, unless otherwise clearly specified and limited, the first feature may be in direct contact with the first feature or the first feature and the second feature through an intermediary indirect contact. Moreover, "above", "above" and "above" the first feature on the second feature may mean that the first feature is directly above or obliquely above the second feature, or simply means that the first feature is higher in level than the second feature. "Below", "beneath" and "beneath" the first feature may mean that the first feature is directly below or obliquely below the second feature, or simply means that the first feature is less horizontally than the second feature.

Referring to Figures 1 to 6, the culture dish of the present utility model can realize the cultivation of microbial cells on both sides of the solid medium, can realize the efficient and selective separation and screening of functional microorganisms with antibacterial activity, and can prepare the separation and cultivation of "simulated in situ environment" It has made the research on the non-contact interaction between microorganisms and the interaction between microorganisms and the environment feasible, and has the advantages of simplicity, high efficiency, and low cost.

A double-sided solid petri dish, which includes: a buckle upper cover 3, a buckle lower cover 4 and a straight dish wall 7, wherein the straight dish wall 7 is arranged between the upper cover and the lower cover, wherein the straight dish A gauze-type permeable membrane bottom 5 with a horizontal structure is arranged inside the wall 7. The upper and lower sides of the gauze-type permeable membrane bottom 5 can be used for cultivating microorganisms. .

As an optional implementation, in some embodiments, the gauze-type permeable membrane bottom 5 has a grid-like fixing structure, and several "T"-shaped column structures 6 are provided on the upper and lower sides of the grid-like fixing structure, "T" The columnar structure 6 is perpendicular to the bottom 5 of the gauze type permeable membrane.

As an optional implementation, in some embodiments, the "T"-shaped column structure 6 includes: a column vertically connected to the gauze-type permeable membrane bottom 5, and a cross bar vertically connected to the end of the column or flat.

As an optional implementation, in some embodiments, the gauze-type permeable membrane bottom 5 is supported by a permeable membrane and a grid-like fixing structure, and the "T"-shaped column structure 6 is arranged at the node of the grid-like fixing structure or on the grid.

As an optional implementation, in some embodiments, the inner wall of the straight vessel wall 7 is provided with a circle of ring-shaped protrusion structures 8 above and below the contact position of the gauze-type permeable membrane bottom 5 .

As an optional implementation, in some embodiments, a plurality of spherical protruding points 10 are provided on the inner wall of the mouth edge of the upper cover and the lower cover, and a circle of spherical protruding rings is provided outside the upper and lower mouth edges of the straight vessel wall 7 11. The spherical protruding point 10 cooperates with the spherical protruding ring 11 to form a buckle structure.

As an optional implementation, in some embodiments, the gauze-type permeable membrane bottom 5 is arranged in the middle of the straight vessel wall 7 and the gauze-type permeable membrane bottom 5 and the straight vessel wall 7 are integrally arranged.

As an optional embodiment, in some embodiments, a quantitative scale 9 is provided on the outer wall of the straight tube wall 7, the quantitative scale 9 is perpendicular to the gauze type permeable membrane bottom 5, and the zero scale of the quantitative scale 9 is located at the quantitative The middle position of the ruler 9 is aligned with the bottom 5 of the gauze type transparent membrane.

As an optional implementation, in some embodiments, the inner diameters of the upper cover and the lower cover are greater than the outer diameter of the straight vessel wall 7 and the difference between the two diameters is kept within a set range.

As an optional implementation, in some embodiments, there is a ventilation gap 12 between the upper cover and the lower cover and the straight vessel wall 7 respectively.

As shown in Figures 2-4, in a complete embodiment, a novel double-sided solid Petri dish and its application can include:

The novel double-sided solid culture dish of the present utility model comprises an upper cover 3, a lower cover 4, a columnar straight tube wall 7, and a gauze-type transparent film bottom 5 at the middle position of the columnar straight tube wall 7; A "T"-shaped column structure 6 is set at the cross point of the lattice support structure of the membrane bottom 5 and on the grid line; a ring-shaped protrusion structure is respectively arranged on the upper and lower inner walls of the connection between the gauze-type transparent membrane bottom 5 and the columnar straight vessel wall 7 8. The annular protrusion structure 8 is perpendicular to the inner wall of the straight cylinder. Spherical protruding points 10 are arranged at the same level inside the mouth edge of the upper and lower covers; spherical protruding rings 11 are arranged on the outer side of the cylindrical straight vessel wall and shallower than the spherical protruding points 10 of the inner wall of the cover; the gauze-type transparent membrane bottom 5 and the upper and lower covers Culture grooves are respectively formed between them. The "T" column structure 6 on the gauze-type permeable membrane bottom 5 forms a joint force with the circular protrusion structure 8 on the inner side of the columnar straight vessel wall 7 to support the solid culture medium closely against the gauze-type permeable membrane bottom 5 does not break away from; The inner diameter of upper cover 3 and lower cover 4 is slightly larger than the outer diameter of columnar straight tube dish 7.

The specific use process is as follows:

1) Complete the sterilization process of the microbial solid medium, place it on a sterile operating table, shake it well, and cool it to about 65 degrees Celsius;

2) Remove the upper cover of the sterile double-sided solid culture dish, pour the sterilized medium cooled to about 65 degrees Celsius on the upper gauze-type permeable membrane bottom plate, and submerge the top of the "T" column structure, cover Put the lid on and wait for the culture medium to solidify;

3) After the upper medium is solidified, invert the double-sided solid culture dish, open the lower cover, pour in the medium cooled to about 65 degrees Celsius, and submerge the top of the "T"-shaped column structure, cover the upper and lower covers, and wait for it to cool down. solidification.

4) After the solidified culture medium is left for a period of time, inoculate microorganisms on both sides or coat other matrix experimental samples.

5) Taking the selective separation of microbial resources that produce active substances as an example, the medium prepared by this double-sided solid culture dish can be coated with an environmental sample with a moderate dilution on one side, and inoculated with indicator bacteria on the other side. Observe the mode of the inhibition zone of the indicator bacteria, and screen out a single colony that produces active substances against the indicator bacteria among the many wild colonies on the other side of the isolated petri dish. Screening in one step, accurate and efficient, shortens time and saves cost, greatly omits the traditional steps of strain isolation and purification, fermentation extract (or agar block method), and then transfers to experiments such as plate observation with indicator bacteria The process greatly improves the efficiency of microbial screening.

The difference from existing technology is:

On the upper and lower covers of the culture dish, there are multiple spherical bumps on the same horizontal line on the inner wall of the upper and lower lids, which are used to form a buckle structure with a spherical protruding ring outside the wall of the columnar straight cylinder, which not only ensures the air inside and outside the dish. Exchange, can avoid the free fall of dish cover again.

Gauze-type permeable membrane bottom, the gauze-type permeable membrane bottom of the dish is in the middle of the straight vessel wall; the gauze-type permeable membrane bottom is permeable, realizing the free exchange of substances between the two sides of the gauze-type permeable membrane bottom ;The permeability of the gauze-type permeable membrane is based on the culture medium above 65 degrees Celsius without obvious leakage; The "T" column structure is perpendicular to the bottom plane of the gauze-type permeable membrane. The "T" column structure is used to fix the poured culture medium and it will not separate from the bottom of the gauze-type permeable membrane even if it is inverted.

Columnar straight dish wall, the columnar straight dish wall is about twice or more the depth of a normal single-layer petri dish, there is a quantitative scale on the outer wall of the dish wall, and the zero scale is located in the middle of the scale and aligned with the bottom of the gauze-type permeable membrane. It is used to measure the thickness of the culture medium; there is a spherical protrusion structure on the outer side of the upper and lower mouth edges of the columnar straight vessel wall, which cooperates with the spherical protrusions on the inner side of the cover to form a buckle structure. The connecting part of the bottom of the gauze-type permeable membrane on the inner side of the wall edge of the columnar straight cylinder has a ring protrusion structure on the upper and lower sides, parallel to the plane of the gauze-type permeable membrane bottom, perpendicular to the inner wall of the straight cylinder wall, and connected to the gauze-type permeable membrane bottom " The upper edge of the T”-shaped column structure is on the same horizontal plane, and the combined action makes the solidified solid medium matrix unable to separate from the gauze-type permeable membrane bottom plate even if it is inverted.

When in use, remove the upper cover, pour the sterilized culture medium into the upper gauze-type permeable membrane bottom plate, and submerge the top of the "T" column structure, cover the upper cover, and wait for the medium to solidify and stabilize Finally, reverse the up and down direction of the dish, open the lower cover, pour the medium in the molten state, and submerge the top of the "T" column structure, cover the upper and lower covers, and wait for it to solidify. After solidification, the "T"-shaped column structure on the bottom of the permeable membrane and the ring-shaped platform structure on the inner wall of the straight cylinder form a joint force to support the medium together even if it is inverted, so that it can be coated on both sides. Microbial or other matrix experimental samples.

The new double-sided solid culture dish of the utility model provides a brand-new way for the pure cultivation and separation of microorganisms with specific functions. The culture medium configured by the double-sided solid culture dish can realize efficient and accurate microbial resources for producing active substances and other functions. Directed selective separation and screening, to achieve one-step screening, accurate and efficient, which not only shortens the time, but also saves costs; makes the non-contact interaction research between microorganisms or between microorganisms and substrates easy and fast, and is designed for uncultured microorganisms The "in situ simulation culture" program provides the possibility.

The utility model has the characteristics of simple and smart design, low cost, convenience and quickness, strong practicability, diverse functions, etc., and is conducive to promoting the research and development in the field of natural microbial resource mining.

In the description of this specification, descriptions referring to the terms "one embodiment", "some embodiments", "example", "specific examples", or "some examples" mean that specific features described in connection with the embodiment or example , structures, materials or features are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the described specific features, structures, materials or characteristics may be combined in any suitable manner in any one or more embodiments or examples. In addition, those skilled in the art can combine and combine different embodiments or examples and features of different embodiments or examples described in this specification without conflicting with each other.

The above-mentioned embodiments are only to illustrate the technical concept and characteristics of the present utility model, and its purpose is to allow those of ordinary skill in the art to understand the content of the present utility model and implement it accordingly, and cannot limit the protection scope of the present utility model with this . All equivalent changes or modifications made according to the essence of the content of the utility model shall fall within the scope of protection of the utility model.

Claims

A double-sided solid culture dish is characterized in that it comprises:

cover;

lower lid;

A straight vessel wall, which is arranged between the upper cover and the lower cover, wherein a gauze-type transparent membrane bottom with a horizontal structure is arranged inside the straight vessel wall, and the upper and lower sides of the gauze-type transparent membrane bottom are Both can be used for cultivating microorganisms, and the upper cover and the lower cover are correspondingly fastened to the upper and lower end surfaces of the straight vessel wall.
The double-sided solid culture dish according to claim 1, wherein the bottom of the gauze-type permeable membrane has a grid-like fixed structure, and the upper and lower sides of the grid-like fixed structure are respectively provided with several "T"-shaped column structures. The "T" column structure is perpendicular to the bottom of the gauze type permeable membrane.
The double-sided solid culture dish according to claim 2, wherein the "T"-shaped column structure comprises: a column vertically connected to the bottom of the gauze-type permeable membrane, and a column vertically connected to the bottom of the column end of the bar or plane.
The double-sided solid culture dish according to claim 3, wherein the bottom of the gauze-type permeable membrane is supported by a permeable membrane and a grid-shaped fixed structure, and the "T"-shaped column structure is arranged on the grid-shaped Fixed structure nodes or grids.
The double-sided solid Petri dish according to claim 1, wherein the inner wall of the straight dish is provided with a circle of protrusions above and below the contact position of the gauze-type permeable membrane bottom.
The double-sided solid Petri dish according to claim 1, wherein the inner walls of the upper cover and the lower cover are provided with a plurality of spherical protruding points, and the outer sides of the upper and lower mouth edges of the straight vessel wall are provided with There is a circle of spherical protruding rings, and the spherical protruding points cooperate with the spherical protruding rings to form a buckle structure.
The double-sided solid culture dish according to claim 1, wherein the bottom of the gauze-type permeable membrane is arranged in the middle of the wall of the straight vessel and the bottom of the gauze-type permeable membrane is in contact with the wall of the straight vessel. All in one setting.
The double-sided solid culture dish according to claim 1, wherein a quantitative scale is provided on the outer wall of the columnar straight tube wall, the quantitative scale is perpendicular to the bottom of the gauze type permeable membrane, and the quantitative scale is vertical to the bottom of the gauze type permeable membrane. The zero scale is located in the middle of the quantitative scale and aligned with the bottom of the gauze-type permeable membrane.
The double-sided solid Petri dish according to claim 1, wherein the inner diameter of the upper cover and the lower cover is greater than the outer diameter of the straight vessel wall and the difference between the diameters of the two remains within a set range Inside.
The double-sided solid Petri dish according to claim 1, characterized in that there are ventilation gaps between the upper cover and the lower cover and the wall of the straight vessel respectively.