阅读说明：本技术 一种可控制微生物培养环境湿度的培养皿及其应用 (Culture dish capable of controlling humidity of microorganism culture environment and application thereof ) 是由 王文琳 应南娇 张曼莉 陈逸璇 于 2021-06-18 设计创作，主要内容包括：本发明涉及微生物培养技术领域,公开了一种可控制微生物培养环境湿度的培养皿及其应用。该培养皿包括相互匹配的皿身和皿盖；所述皿身的底部设有干燥剂；所述皿身的顶部设有密封下层,所述密封下层与皿盖的顶部紧密贴合；所述皿盖的顶面上非圆心位置设有加样通孔一,所述密封下层设有加样通孔二,皿盖与皿身相对旋转后所述加样通孔一与加样通孔二的位置重合。本发明的培养皿利用干燥剂作为微生物培养环境,并采用特殊的密封结构,能够调节微生物培养环境湿度,并控制培养过程中环境湿度相对恒定,因而能用于研究环境湿度或干燥条件对微生物生长繁殖的影响,实现一定湿度下微生物的培养。(The invention relates to the technical field of microbial culture, and discloses a culture dish capable of controlling the humidity of a microbial culture environment and application thereof. The culture dish comprises a dish body and a dish cover which are matched with each other; a drying agent is arranged at the bottom of the dish body; the top of the dish body is provided with a lower sealing layer, and the lower sealing layer is tightly attached to the top of the dish cover; and a first sample adding through hole is formed in the non-circle center position on the top surface of the dish cover, a second sample adding through hole is formed in the lower sealed layer, and the first sample adding through hole and the second sample adding through hole coincide in position after the dish cover and the dish body rotate relatively. The culture dish provided by the invention utilizes the drying agent as a microorganism culture environment, adopts a special sealing structure, can adjust the humidity of the microorganism culture environment, and controls the relative constancy of the environmental humidity in the culture process, so that the culture dish can be used for researching the influence of the environmental humidity or the drying condition on the growth and the propagation of microorganisms, and realizes the culture of the microorganisms under a certain humidity.)

1. A culture dish capable of controlling the humidity of a microorganism culture environment comprises a dish body (1) and a dish cover (2) which are matched with each other, and is characterized in that a drying agent is arranged at the bottom of the dish body (1); the top of the dish body (1) is provided with a lower sealing layer (3), and the lower sealing layer (3) is tightly attached to the top of the dish cover (2); and a non-circle center position on the top surface of the dish cover (2) is provided with a first sample adding through hole (4), the sealed lower layer (3) is provided with a second sample adding through hole (5), and the dish cover (2) and the dish body (1) rotate relatively, and then the positions of the first sample adding through hole (4) and the second sample adding through hole (5) are coincided.

2. A culture dish according to claim 1, wherein the dish body (1) is provided with a filter layer (6) having a diameter equal to the inner diameter of the dish body (1); the filter layer (6) is provided with a plurality of through holes (7).

3. The culture dish according to claim 2, wherein the first sample adding through hole (4) is arranged on the top surface of the dish cover (2) close to the circle center; the aperture of the through hole (7) is sequentially increased from the circle center of the filter layer (6) to the outside.

4. The culture dish according to claim 3, wherein the through holes (7) are arranged in at least two layers of through hole rings from inside to outside along the circumferential direction of the filter layer (6), the hole diameters of the through holes (7) in two adjacent layers of through hole rings are different by 1-2 mm, and the hole diameter of the through hole in the innermost layer of through hole ring is 2.7-3.3 mm.

5. A culture dish according to claim 2, wherein the distance between the filter layer (6) and the lower sealing layer (3) is 1/4-1/3 of the height of the dish body (1).

6. The culture dish according to claim 1, wherein the dish body (1) is in threaded connection with the dish cover (2), and the lower sealing layer (3) is tightly attached to the top of the dish cover (2) after screwing.

7. The culture dish of claim 1, wherein the desiccant is silica gel or flour.

8. A method for cultivating microorganisms using a culture dish according to any of claims 2 to 7, comprising the steps of:

(1) rotating the dish cover to enable the position of the sample adding through hole I on the dish cover to coincide with the position of the sample adding through hole II on the dish body;

(2) adding a sample into the dish body through the first sample adding through hole and the second sample adding through hole, wherein the sample comprises microorganisms and water or comprises microorganisms, water and nutrient substances; controlling the water content of the drying agent by controlling the added sample amount;

(3) rotating the dish cover to stagger the positions of the first sample adding through hole and the second sample adding through hole, and tightly attaching the lower sealing layer to the top of the dish cover to culture microorganisms.

9. The method of claim 8, wherein in step (2), the relationship between the amount of sample and the humidity of the culture environment is as follows:

wherein, W_gWater content of desiccant, W₁Is the mass of the desiccant, W₂Is the sample mass.

10. The method of claim 8, wherein in step (2), the sample is added to the non-through-hole region of the filter layer.

Technical Field

The invention relates to the technical field of microbial culture, in particular to a culture dish capable of controlling the humidity of a microbial culture environment and application thereof.

Background

Microorganisms are microorganisms which cannot be observed by naked eyes, and comprise bacteria, viruses, fungi, some small protists, microscopic algae and the like, so that the microorganisms cover various beneficial and harmful types and are widely applied to the fields of food, medicine, industry and agriculture, environmental protection, sports and the like.

A petri dish is a laboratory vessel for the cultivation of microorganisms or cells, consisting of a flat disc-shaped bottom and a lid, generally made of glass or plastic. The culture dish is mainly made of plastic and glass, and the glass culture dish can be used for culturing plant materials and microorganisms and for adherent culture of animal cells; the plastic culture dish mainly adopts polyethylene materials, and comprises disposable and reusable materials, is suitable for operations of inoculation, streaking and bacteria separation in a laboratory, and can also be used for culture of plant materials.

In the process of culturing the microorganisms, the environmental humidity can have important influence on the growth and the propagation of the microorganisms, and the environmental humidity required for culturing different microorganisms is different. However, the culture dish used in the laboratory at present is generally in a structure consisting of a plane disc-shaped bottom and a cover, so that the humidity of the microorganism culture environment is difficult to adjust, and the relative constancy of the humidity of the environment cannot be controlled in the culture process, so that the culture dish cannot be used for researching the influence of the humidity of the environment or the drying condition on the growth of microorganisms, and cannot regulate the growth of the microorganisms by controlling the humidity.

Disclosure of Invention

In order to solve the technical problems, the invention provides a culture dish capable of controlling the humidity of a microorganism culture environment and application thereof. The culture dish utilizes a drying agent as a microorganism culture environment, adopts a special sealing structure, can adjust the humidity of the microorganism culture environment, and controls the relative constancy of the environmental humidity in the culture process.

The specific technical scheme of the invention is as follows:

a culture dish capable of controlling the humidity of a microorganism culture environment comprises a dish body and a dish cover which are matched with each other; a drying agent is arranged at the bottom of the dish body; the top of the dish body is provided with a lower sealing layer, and the lower sealing layer is tightly attached to the top of the dish cover; and a first sample adding through hole is formed in the non-circle center position on the top surface of the dish cover, a second sample adding through hole is formed in the lower sealed layer, and the first sample adding through hole and the second sample adding through hole coincide in position after the dish cover and the dish body rotate relatively.

A method of culturing microorganisms using the culture dish, comprising the steps of:

(1) rotating the dish cover to enable the position of the sample adding through hole I on the dish cover to coincide with the position of the sample adding through hole II on the dish body;

(2) adding a sample into the dish body through the first sample adding through hole and the second sample adding through hole, wherein the sample comprises microorganisms and water or comprises microorganisms, water and nutrient substances; controlling the water content of the drying agent by controlling the added sample amount;

(3) rotating the dish cover to stagger the positions of the first sample adding through hole and the second sample adding through hole, and tightly attaching the lower sealing layer to the top of the dish cover to culture microorganisms.

In the culture dish, microorganisms are adsorbed in the drying agent and grow and reproduce in pores of the drying agent, and the drying agent is used as a culture environment, so that the humidity of the microorganism culture environment can be controlled by controlling the water content in the drying agent. When a sample is added, water in the sample is also adsorbed into the drying agent, and the water content in the drying agent, namely the humidity of the microbial culture environment, can be controlled by controlling the amount of the added sample; utilize the cooperation between ware lid, sealed lower floor and application of sample through-hole one and application of sample through-hole two, rotatory ware lid can realize sealed after adding the sample, prevents during the moisture evaporation in the drier or the moisture in the environment is adsorbed the drier to control microorganism and cultivate environment humidity constancy relatively.

When the added sample does not contain nutrient substances, the method can be used for researching the influence of the environmental humidity or the drying condition on the growth and the propagation of the microorganisms; when the added sample contains nutrient substances, the culture of the microorganism under certain humidity can be realized.

Preferably, a filter layer with the diameter the same as the inner diameter of the dish body is arranged in the dish body; the filter layer is provided with a plurality of through holes.

The filter layer is arranged to have the following functions: when the bacteria liquid is added to the non-through hole position on the filter layer, the filter layer can enable the bacteria liquid to be leveled to a certain degree, and then the bacteria liquid is uniformly adsorbed into the drying agent at the bottom of the dish body, so that the humidity in the drying agent can be controlled to be uniform, and the microorganisms can be controlled to be uniformly distributed in the drying agent.

Furthermore, the first sample adding through hole is arranged on the top surface of the dish cover and close to the circle center; the aperture of the through hole is sequentially increased from the circle center of the filter layer to the outside.

Because application of sample through-hole one is located the position that is close to the centre of a circle on the dish lid top surface, consequently, when adding the sample in to the culture dish through application of sample through-hole one and application of sample through-hole two, the sample can concentrate the distribution in the position that is close to the centre of a circle, leads to humidity and microorganism to distribute inhomogeneous in the drier (humidity is higher than the marginal drier humidity of culture dish in the drier of being close to culture dish central point, and the microorganism distributes also more). In order to solve the problems, the through holes on the filter layer are designed into structures which are sequentially enlarged from the circle center to the outside, so that a sample can flow on the filter layer to a position far away from the circle center, and the humidity and microorganisms in the drying agent are uniformly distributed.

Furthermore, the through holes are arranged into at least two layers of through hole rings from inside to outside along the circumferential direction of the filter layer, the aperture difference of the through holes in the two adjacent layers of through hole rings is 1-2 mm, and the aperture of the through hole in the innermost layer of through hole ring is 2.7-3.3 mm.

Under the condition that the pore diameters of the through holes in the innermost layer are the same, the pore diameters of the through holes in the two adjacent layers of through hole rings are too different or too small, so that a sample cannot uniformly enter a drying agent, and the humidity and microorganism distribution in the drying agent is not uniform, specifically: when the pore size difference is too small, the downward flowing speed of the sample through the filter layer is too low, so that the sample is easy to accumulate on the filter layer, further the downward flowing of the sample at a position close to the center of the circle (namely the position where the sample is added) is more, the humidity of the drying agent at a position close to the center of the culture dish is higher, and the distribution of microorganisms is more; when the pore size difference is too large, the downward flow speed of the sample through the filter layer is too high, so that the sample cannot flow to the edge of the filter layer, and the desiccant near the center of the culture dish has high humidity and more microorganism distribution.

Further, the distance between the filter layer and the lower sealing layer is 1/4-1/3 of the dish height.

Preferably, the dish body is in threaded connection with the dish cover, and the lower sealing layer is tightly attached to the top of the dish cover after the dish body is screwed.

Preferably, the drying agent is silica gel or flour.

Silica gel and flour do not adversely affect the growth and proliferation of microorganisms and therefore can be used as a desiccant in the present invention.

Preferably, in step (2), the relationship between the amount of sample and the humidity of the culture environment is as follows:

wherein, W_gWater content of desiccant, W₁Is the mass of the desiccant, W₂Is the sample mass.

The sample has a low level of microorganisms, so that when the moisture content of the desiccant is calculated, the mass of the sample can be used to approximate the mass of water in the sample, i.e., the mass of water adsorbed by the desiccant after the sample is added.

Preferably, in step (2), the sample is added to the non-through-hole region of the filter layer.

Compared with the prior art, the invention has the following advantages:

(1) the culture dish provided by the invention utilizes the drying agent as a microorganism culture environment, adopts a special sealing structure, can adjust the humidity of the microorganism culture environment, and controls the relative constancy of the environmental humidity in the culture process, so that the culture dish can be used for researching the influence of the environmental humidity or the drying condition on the growth and the propagation of microorganisms, and realizes the culture of the microorganisms under a certain humidity;

(2) through the design of the filter layer and the through holes in the filter layer, the sample can uniformly flow to the bottom of the dish body and is uniformly mixed with the drying agent, so that the humidity in the drying agent can be controlled to be uniform, and the microorganisms can be controlled to be uniformly distributed in the drying agent.

Drawings

FIG. 1 is a schematic view of a structure of a culture dish according to the present invention;

FIG. 2 is a top view of the lower seal layer of the present invention;

FIG. 3 is a top view of the capsule of the present invention;

FIG. 4 is a top view of the combination of the lower seal layer and capsule of the present invention;

FIG. 5 is a top view of a filter layer according to the present invention.

The reference signs are: vessel 1, vessel lid 2, sealed lower floor 3, application of sample through-hole 4, application of sample through-hole two 5, filter layer 6, through-hole 7.

Detailed Description

The present invention will be further described with reference to the following examples.

In an application example, the method for calculating the sample amount according to the set moisture content of the drying agent is as follows:

wherein, W_gWater content of desiccant, W₁Is the mass of the desiccant, W₂Is the sample mass.

Example 1

A culture dish capable of controlling the humidity of a microorganism culture environment comprises a dish body 1 and a dish cover 2 which are matched with each other, wherein the dish body 1 is in threaded connection with the dish cover 2. Silica gel is uniformly distributed at the bottom of the dish body 1.

The top of ware body 1 has set firmly sealed lower floor 3, and the back of screwing between ware body 1 and the ware lid 2, sealed lower floor 3 closely laminates with the top of ware lid 2. As shown in fig. 2 and 3, a first sample adding through hole 4 is arranged on the top surface of the dish cover 2 near the circle center, a second sample adding through hole 5 with the same aperture as the first sample adding through hole 4 is arranged on the lower sealing layer 3, and the positions of the first sample adding through hole 4 and the second sample adding through hole 5 are overlapped after the dish cover 2 and the dish body 1 rotate relatively.

The filter layer 6 with the same diameter as the inner diameter of the dish body 1 is fixedly arranged in the dish body 1, and the distance between the filter layer 6 and the lower sealing layer 3 is 1/4 of the height of the dish body 1. Four layers of through hole rings are arranged on the filter layer 6 from inside to outside along the circumferential direction, each layer of through hole ring comprises six through holes 7, the aperture difference of the through holes 7 in the two adjacent layers of through hole rings is 1mm, and the aperture of the through hole in the innermost layer of through hole ring is 3 mm.

Example 2

This example is different from example 1 in that silica gel was replaced with flour in this example.

Example 3

The difference between this embodiment and embodiment 2 is that in this embodiment, the distance between the filter layer 6 and the lower sealing layer 3 is 1/3 of the height of the dish body 1; the aperture difference of the through holes 7 in the two adjacent layers of through hole rings is 2mm, and the aperture of the through hole in the innermost layer of through hole ring is 2.7 mm.

Example 4

The difference between this embodiment and embodiment 1 is that in this embodiment, the sealing lower layer 3 and the filter layer 6 are detachably connected with the dish body 1.

In this embodiment, after the sealing lower layer 3 and the filter layer 6 are taken out, a drying agent can be added to or taken out from the dish body 1, thereby realizing the recycling of the culture dish.

Comparative example 1

The present comparative example is different from example 1 in that the aperture of the through-hole 7 in the adjacent two-layer through-hole ring differs by 0.5mm in the present comparative example.

Comparative example 2

The present comparative example is different from example 3 in that the aperture of the through-hole 7 in the adjacent two-layer through-hole ring differs by 2.5mm in the present comparative example.

Application example 1

The culture dish of example 1 was used for culturing microorganisms, the mass of silica gel in the culture dish was 5g, and the microorganism culture method was as follows:

(1) mixing Escherichia coli with PBS buffer solution to obtain sample with Escherichia coli content of 10⁹CFU/mL, sample density 1.66 g/mL;

(2) setting the water content of the drying agent to be 20%, and calculating the volume of a sample needing to be added to be 1.250 mL;

(3) rotating the dish cover 2 to ensure that the position of the sample adding through hole I4 on the dish cover 2 is superposed with the position of the sample adding through hole II 5 on the dish body 1;

(4) adding 1.250mL of sample into the vessel body 1 through the first sample adding through hole 4 and the second sample adding through hole 5 by using a pipette, and controlling the sample to be added into the non-through hole area of the filter layer;

(5) rotating the dish cover 2 to stagger the positions of the first sample adding through hole 4 and the second sample adding through hole 5, and tightly attaching the lower sealed layer 3 to the top of the dish cover 2, after the sample flows into the bottom of the dish body 1, putting the culture dish into a 30 ℃ incubator to culture the microorganisms.

After 48h of culture, the moisture content in the desiccant measured by a hygrometer is 20.3%, and the error between the moisture content and the set moisture content is 0.3%, which shows that the culture dish can control the humidity of the microorganism culture environment to be relatively constant; the moisture content of the desiccant in the center of the culture dish is measured to be 20.5% by a hygrometer, the moisture content of the desiccant at the edge of the culture dish is measured to be 19.1%, and the difference between the moisture content of the desiccant and the moisture content of the desiccant is small, so that the humidity in the desiccant is uniform in the culture dish.

Application example 2

The culture dish of example 1 was used for culturing microorganisms, the mass of silica gel in the culture dish was 5g, and the microorganism culture method was as follows: (1) mixing Escherichia coli with PBS buffer solution to obtain sample with Escherichia coli content of 10⁹CFU/mL, sample density 1.66 g/mL;

(2) setting the water content of the drying agent to be 15%, and calculating the volume of a sample needing to be added to be 0.882 mL;

(3) rotating the dish cover 2 to ensure that the position of the sample adding through hole I4 on the dish cover 2 is superposed with the position of the sample adding through hole II 5 on the dish body 1;

(4) adding 0.882mL of sample into the vessel body 1 through the first sample adding through hole 4 and the second sample adding through hole 5 by using a pipette, and controlling the sample to be added into the non-through hole area of the filter layer;

(5) rotating the dish cover 2 to stagger the positions of the first sample adding through hole 4 and the second sample adding through hole 5, and tightly attaching the lower sealed layer 3 to the top of the dish cover 2, after the sample flows into the bottom of the dish body 1, putting the culture dish into a 30 ℃ incubator to culture the microorganisms.

After 48h of culture, the moisture content in the desiccant measured by a hygrometer was 15.1%, and the error from the set moisture content was 0.1%, indicating that the culture dish of the present invention can control the humidity of the microorganism culture environment to be relatively constant.

Application example 3

The culture dish of example 2 was used for culturing microorganisms, the mass of flour in the culture dish was 5g, and the microorganism culture method was as follows:

(1) mixing Escherichia coli with PBS buffer solution to obtain sample with Escherichia coli content of 10⁹CFU/mL, sample density 1.66 g/mL;

(2) setting the water content of the drying agent to be 20%, and calculating the volume of a sample needing to be added to be 1.250 mL;

(3) rotating the dish cover 2 to ensure that the position of the sample adding through hole I4 on the dish cover 2 is superposed with the position of the sample adding through hole II 5 on the dish body 1;

(4) adding 1.250mL of sample into the vessel body 1 through the first sample adding through hole 4 and the second sample adding through hole 5 by using a pipette, and controlling the sample to be added into the non-through hole area of the filter layer;

(5) rotating the dish cover 2 to stagger the positions of the first sample adding through hole 4 and the second sample adding through hole 5, and tightly attaching the lower sealed layer 3 to the top of the dish cover 2, after the sample flows into the bottom of the dish body 1, putting the culture dish into a 30 ℃ incubator to culture the microorganisms.

After 48h of culture, the moisture content in the desiccant measured by a hygrometer was 19.2%, and the error from the set moisture content was 0.8%, indicating that the culture dish of the present invention can control the humidity of the microorganism culture environment to be relatively constant.

Application example 4

The culture dish of example 3 was used for culturing microorganisms, the mass of flour in the culture dish was 5g, and the microorganism culture method was as follows:

(1) mixing Escherichia coli with liquid TGY culture medium to obtain sample with Escherichia coli content of 10⁹CFU/mL, sample density 1.66 g/mL;

(2) setting the water content of the drying agent to be 20%, and calculating the volume of a sample needing to be added to be 1.250 mL;

(3) rotating the dish cover 2 to ensure that the position of the sample adding through hole I4 on the dish cover 2 is superposed with the position of the sample adding through hole II 5 on the dish body 1;

(4) adding 1.250mL of sample into the vessel body 1 through the first sample adding through hole 4 and the second sample adding through hole 5 by using a pipette, and controlling the sample to be added into the non-through hole area of the filter layer;

(5) rotating the dish cover 2 to stagger the positions of the first sample adding through hole 4 and the second sample adding through hole 5, and tightly attaching the lower sealed layer 3 to the top of the dish cover 2, after the sample flows into the bottom of the dish body 1, putting the culture dish into a 30 ℃ incubator to culture the microorganisms.

After 48h of culture, the moisture content in the desiccant measured by a hygrometer is 19.0%, and the error between the moisture content and the set moisture content is 1.0%, which shows that the culture dish can control the humidity of the microorganism culture environment to be relatively constant; the moisture content of the desiccant in the center of the culture dish is measured to be 20.6% by a hygrometer, the moisture content of the desiccant at the edge of the culture dish is measured to be 19.1%, and the difference between the moisture content of the desiccant and the moisture content of the desiccant is small, so that the humidity in the desiccant is uniform in the culture dish.

Application example 5

The culture dish of comparative example 1 was used to culture microorganisms, the mass of silica gel in the culture dish was 5g, and the method of culturing microorganisms was the same as in application example 1.

After 48h of culture, the moisture content of the desiccant in the center of the culture dish is 23.3% and the moisture content of the desiccant at the edge is 18.1% as measured by a hygrometer, which is 5.2% different from that of the desiccant in application example 1, indicating that the difference between the pore diameters of the through holes in the two adjacent layers of through hole rings is too small, which results in uneven moisture distribution in the desiccant (the humidity of the desiccant near the center of the culture dish is higher than that of the desiccant at the edge of the culture dish).

Application example 6

The culture dish of comparative example 2 was used for culturing microorganisms, the mass of flour in the culture dish was 5g, and the method of culturing microorganisms was the same as in application example 4.

After 48h of culture, the moisture content of the desiccant in the center of the culture dish is measured to be 24.7% by a hygrometer, the moisture content of the desiccant at the edge is measured to be 17.5%, the difference is 7.2%, and is obviously higher than that in application example 4, which shows that the aperture difference of the through holes in the two adjacent layers of through hole rings is too large, so that the moisture distribution in the desiccant is uneven (the humidity of the desiccant at the position close to the center of the culture dish is higher than that of the desiccant at the edge of the culture dish).

The raw materials and equipment used in the invention are common raw materials and equipment in the field if not specified; the methods used in the present invention are conventional in the art unless otherwise specified.

The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and all simple modifications, alterations and equivalents of the above embodiments according to the technical spirit of the present invention are still within the protection scope of the technical solution of the present invention.