JP2678293B2 - Fungal culture and observation container - Google Patents

Description

TECHNICAL FIELD The present invention relates to a container used for culturing a fungus and observing the cultivated fungus, and in particular, a state in which the fungus is cultivated in the container. As it is, with a macroscopic observation from both front and back sides of the giant colony (giant colony),
The present invention relates to a fungal culture / observation container that can be used together with observation (microscopic examination) of morphological characteristics of fungal elements such as hyphae and spores.

[Conventional technology]

When the pathogenic fungus isolated from a patient is to be identified bacteriologically, in general, a specimen is embedded in an appropriate medium contained in a container and inoculated, and the fungus is cultured in the container for, for example, about 1 to 2 weeks. To grow a huge colony, and visually observe the growth rate of the huge colony, the state of growth, each color of the front and back of the colony, the presence or absence of hyphae and wrinkles, and for morphological identification, Take out a part of the cultured fungus from the container and place it on a slide glass, drop an appropriate amount of a staining solution such as lactophenol cotton blue liquor on it, cover it with a cover glass to make a stained specimen, By microscopic examination, the morphological characteristics of fungal elements such as hyphae and spores were also observed. Alternatively, while performing the above-mentioned macroscopic observation, separately, by a slide glass culture method, that is, a medium piece is placed on a slide glass, the medium piece is inoculated with a fungus, and a cover glass is placed on the upper surface of the medium. Then, the fungus was cultivated in a Petri dish, and after culturing, a stained specimen of the fungal element adhered to the cover glass was prepared and microscopically examined to observe the morphological characteristics of the fungal element.

[Problems to be solved by the invention]

In the conventional inspection method described above, after culturing the fungus in the container and performing macroscopic observation of this huge settlement, when performing a microscopic examination, a part of the cultured fungus is taken out of the container and stained. Or, while observing macroscopic colonies with the naked eye, separately culture the fungus by the slide glass culture method, take out the slide glass together with the medium from the Petri dish, and cover the cover glass with sterile tweezers from the top of the medium. It had to be peeled off and used to make a stained specimen of the fungal element attached to the cover glass before microscopic examination. For this reason, the operation for the microscopic examination is very complicated, which requires time and labor. Also, in the process of taking out the fungal element from the container or petri dish and making a precursor sample, the inspector inhales the fungal element that easily floats in the air, such as spores, from the mouth and flowers, and is infected with the pathogenic fungus. There is also a risk of doing.

The present invention has been made in view of the above problems in the conventional inspection method, in which a fungus is cultured in a container, and the fungus is cultured in the container, and the huge colony It is a technical object to provide a fungal culture / observation container capable of comprehensively performing macroscopic observation from both front and back sides and observation (microscopic examination) of morphological characteristics of fungal elements such as hyphae and spores. And

[Means for solving the problem]

The present invention provides a means for achieving the above object,
The fungal culture and observation container is formed of a transparent thin plate material and has a smooth flat surface portion, and in the flat surface portion, a medium storage chamber for storing a solid medium, a moisture storage chamber for storing sterile purified water, The body is also formed with a transparent thin plate-like material and a body part in which a communication groove that connects the water holding chamber and the medium containing chamber and a dyeing solution reservoir that stores the dyeing solution are recessed, and it is in close contact with the flat part of the body part. Then, the medium storage chamber, the water holding chamber,
A through-hole that closes each upper opening surface of the communication groove and the staining solution reservoir, and is rotatably and slidably engaged with the main body, and that at least coincides with the opening position of the culture medium storage chamber with the rotational movement thereof, and Similarly, with the rotation movement, a lid body portion having an injection hole that matches the opening position of the dyeing liquid reservoir, and an adhesive agent is applied to one surface of the lid portion, and the peripheral portion is the peripheral portion of the through hole of the lid body portion. The gist of the present invention is that it is composed of a transparent sticker with an adhesive that is attached and closes the through hole.

[Action]

In the fungal culture / observation container having the above structure, the solid medium is stored in the medium storage chamber of the main body, and the lid is engaged with the main body. When performing a fungal test, the lid part is rotated and slid with respect to the main body part so that the through hole of the lid part matches the opening position of the culture medium storage chamber, and the tip of a platinum wire or the like is passed through the through hole. Is inserted into the medium storage chamber, the sample is embedded in the solid medium stored in the medium storage chamber and inoculated, and then the through hole is closed by a transparent seal with an adhesive. At this time, sterilized purified water is put in the water holding chamber. In this state, for example 1-2 weeks,
Culturing of fungi is carried out by maintaining the container at a predetermined temperature. At this time, since the water storage chamber and the medium storage chamber are communicated with each other via the communication passage formed by the communication groove of the main body and the lower surface of the lid, even if the culture period is long, Moisture prevents the medium from drying out during the culture period. Here, since both the main body and the lid are made of a transparent material, a huge colony of fungi formed by growing in the medium can be visually observed from the front and the back, and its growth can be visually observed. You can check the speed, growth condition, color tone of the community, presence of hyphae and wrinkles. Then, after culturing, it is confirmed that fungal elements such as hyphae and spores are attached to the adhesive-coated surface of the adhesive-attached transparent seal that blocks the through-hole of the lid, or the adhesive of the transparent seal When mycelia, spores, etc. are not attached to the coated surface, lightly press the container with your fingertips to attach the hyphae, spores, etc. to the transparent sticky adhesive coated surface, and then attach the lid to the main body. Rotate it to match the injection hole of the lid with the opening of the dye reservoir in the main unit, inject the dye into the dye reservoir through the injection hole, and then rotate the lid again with respect to the main unit. By moving, the transparent seal is moved to the flat surface of the main body in the vicinity of the dyeing liquid reservoir, and then the container is tilted to flow the dyeing liquid from the dyeing liquid reservoir to the position of the transparent seal. At this time, both the main body and the lid are made of a transparent thin plate material, and the lower surface of the lid is in close contact with the smooth flat surface of the main body. It can be handled, the container is fixed on the stage of the microscope as it is, the objective lens is brought close to the position of the transparent seal, and the hyphae, spores, etc. stained and attached to the adhesive coated surface of the transparent seal at a predetermined magnification Can be observed.

〔Example〕

Hereinafter, preferred embodiments of the present invention will be described with reference to the drawings.

FIGS. 1 and 2 show an embodiment of the present invention.
FIG. 2 is a perspective view showing a state in which a container for fungal culture and observation is separated into its constituent members, and FIG. 2 is a vertical cross-sectional view of the container in use.

This container is composed of a main body portion 10 and a lid portion 12 which are each formed of a thin plate-shaped transparent plastic material. An annular concave portion 14 is formed on the peripheral portion of the main body portion 10, while an annular protruding portion 16 is formed on the peripheral portion of the lid portion 12, and the annular protruding portion 16 is formed on the annular concave portion 14. By fitting, the main body 10 and the lid 12 are integrally engaged. Then, the main body portion 10 has a smooth flat surface portion 18,
When the lid body portion 12 is engaged with the body portion 10, the lower surface of the lid body portion 12 comes into close contact with the flat surface portion 18 thereof. Also, the lid part
12 can rotate and slide while being engaged with the main body 10.

The main body 10 has a flat surface portion 18 on which a medium storage chamber 20, a water holding chamber 22, a communication groove 24 for connecting the medium holding chamber 20 and the water holding chamber 22 and a pair of stain solution reservoirs 26, 26 'are provided. It is recessed. These medium storage chamber 20, water holding chamber 22, communication groove 24
The upper opening surfaces of the dyeing fluid reservoirs 26, 26 'are closed by engaging the lid portion 12 with the body portion 10. At the time of this blockage, as shown in FIG.
A communication passage 28 is formed by 24 and the lower surface of the lid portion 12.
An agar medium 30 is stored in the medium storage chamber 20, and a water absorbing member 32 such as a sponge that absorbs sterilized purified water is stored in the water holding chamber 22. In addition, the stain reservoirs 26, 2
A dye solution is injected into 6 '.

On the other hand, the lid 12 has a pair of through holes 34, 34 'and a small-diameter injection hole 36 formed therein. The through holes 34, 34 ′ and the injection hole 36 are formed by rotating the lid portion 12 with respect to the main body portion 10, so that the through holes 34, 34 ′ are formed in the medium containing chamber 20 and the water holding chamber 22 of the main body portion 10. The injection hole 36 is provided at a predetermined position so as to coincide with each opening position of the dye solution reservoirs 26 and 26 'so as to coincide with each opening position.

In addition, an adhesive agent is applied to one surface and the peripheral edge portion is attached to the peripheral edge portion of the through holes 34, 34 'of the lid portion 12 as shown in FIG. Adhesive transparent seals 38 and 38 'for closing the injection hole 36 and a transparent adhesive seal (not shown) for closing the injection hole 36 are also provided.

Next, FIG. 3 (A), which shows the container as viewed from above.
The procedure for identifying a fungus using this fungal culture / observation container will be described with reference to (E). In the embodiment shown in FIGS. 1 and 2, a pair of through holes 34, 34 'are formed in the lid portion 12 and a pair of stain solution holders 26, 26' are formed in the main body portion 10. Is formed, but this is 1
This is because one container can perform two microscopic examinations, and one through hole and one staining solution reservoir may be provided. In the following description, only one through hole 34 and one staining solution reservoir 26 are provided. It has been formed.

The container accommodates the agar medium 30 in the medium storage chamber 20 of the main body 10, fits the annular protrusion 16 in the annular recess 14, and integrally engages the lid 12 with the main body 10. deep. Also, the lid part
The 12 through holes 34 and the dye solution reservoir 26 are closed by a transparent seal with an adhesive.

When the fungus is inspected with this container, first, the transparent seal that closes the through hole 34 is peeled off from the peripheral portion of the through hole 34, and the lid portion 12 is rotatably slid with respect to the main body portion 10. As shown in FIG. (A), the through hole 34 of the lid 12 is provided in the moisture holding chamber 22.
The sterilized purified water is injected into the water holding chamber 22 through the through hole 34 so that the sterilized purified water is sucked into the water absorbing member 32. Next, the lid portion 12 is rotationally moved in the direction of the arrow a with respect to the main body portion 10, and as shown in FIG.
To the opening position of the medium storage chamber 20, insert the tip of a platinum wire or the like into the medium storage chamber 20 through the through hole 34, and inoculate the agar medium 30 stored in the medium storage chamber 20 by embedding a sample. . After inoculation, as shown in FIG. 3 (C), the through hole 34 is closed by a transparent seal 38 with an adhesive. In this state, the container is kept in an incubator and maintained at room temperature to about 37 ° C., and the fungus is cultured for about 1 to 2 weeks. At this time, since the medium storage chamber 20 communicates with the water holding chamber 22 via the communication passage 28, it is possible to prevent the agar medium from drying during the culture period due to the water in the water holding chamber 22.

During and after this culture period, from the front side and the back side of the container, through the transparent lid portion 12 and the main body portion 10, by observing macroscopic macroscopic colonies of fungi formed by growth on the agar medium, Its growth rate, the state of growth, the color of the village,
Check for mycelia and wrinkles.

Then, when the culture is completed, mycelia on the adhesive coated surface of the transparent sticker 38 with an adhesive that closes the through hole 34 of the lid portion 12,
Make sure that fungal elements such as spores are attached. At this time, when hyphae, spores, etc. are not attached to the adhesive-coated surface of the transparent seal 38, the mycelia, spores, etc. are attached to the adhesive-coated surface of the transparent seal 38 by lightly pressing the container with a fingertip. After that, the transparent seal 40 that closes the injection hole 36 of the lid 12 is peeled off from the periphery of the injection hole 36, and the injection hole 36 of the lid 12 is replaced with the main body 10 as shown in FIG. 3 (D). Staining liquid reservoir
The dye solution is injected into the dye solution reservoir 26 through the injection hole 36 in a state of being aligned with the opening position of 26. Subsequently, the lid portion 12 is rotationally moved in the direction of the arrow b with respect to the main body portion 10, and FIG.
As shown in, by moving the transparent seal 38 to the flat surface of the main body portion 10 in the vicinity of the dyeing liquid reservoir 26, tilting the container causes the dyeing liquid from the dyeing liquid reservoir 26 to be transferred to the transparent seal 38 of the flat surface portion of the main body portion 10. Flow to position. Then, as in the case of a normal stained specimen, the container is placed and fixed on the stage of the microscope as it is, the objective lens is brought close to the position of the transparent seal 38, and adhered to the adhesive coated surface of the transparent seal 38 and stained. Observe the morphological features such as mycelia and spores.

〔The invention's effect〕

Since the present invention is configured and operates as described above, when the fungal culture / observation container according to the present invention is used, the fungus culture is cultivated in the container, and the container is used from both front and back surfaces. Since macroscopic observation and microscopic examination can be performed, the pathogenic fungus can be accurately identified,
In addition, the operation for microscopic examination is greatly simplified and speeded up, and since the microscopic examination can be performed with the fungal element enclosed in the container, the spores and other fungal elements can be removed during the process of making the stained specimen. There is no longer any risk of the inspector inhaling through the mouth or nose to get infected with the pathogenic fungus.

[Brief description of the drawings]

1 and 2 show an embodiment of the present invention, FIG. 1 is a perspective view showing a state in which a fungal culture / observation container is separated into its constituent members, and FIG. 2 is a longitudinal section in the use state of this container. FIGS. 3A to 3E are plan views for explaining a procedure for identifying a fungus using this container. 10 …… Main body part, 12 …… Lid body part, 18 …… Flat part, 20 …… Medium-containing chamber, 22 …… Water holding chamber, 24 …… Communication groove, 26,26 ′ …… Staining solution reservoir, 30 …… Agar medium, 32 …… Water absorbing member, 34,34 ′ …… Transparent hole, 36 …… Injection hole, 38,38 ′ …… Transparent seal with adhesive.

Claims (1)

(57) [Claims] 1. A transparent thin plate-shaped material having a smooth flat surface portion on which a medium storage chamber for storing a solid medium, a moisture holding chamber for holding sterilized purified water, and this moisture holding chamber. And a communication groove that communicates with the culture medium storage chamber, and a main body portion in which a staining solution reservoir for storing a staining solution is recessed, and formed of a transparent thin plate-like material, in close contact with the flat surface portion of the main body portion, The upper opening surfaces of the culture medium storage chamber, the water storage chamber, the communication groove, and the dyeing liquid reservoir are closed and rotatably slidably engaged with the main body portion, and at least the culture medium storage chamber A through-hole that matches the opening position, and a lid part in which an injection hole that matches the opening position of the dyeing liquid reservoir is also formed along with the rotational movement, and an adhesive is applied to one surface,
A fungal culture / observation container comprising a transparent seal having a peripheral edge portion adhered to the peripheral edge portion of the through hole of the lid body and closing the through hole.