Scanning electron microscope sample preparation method suitable for fungi

A technology of sample preparation and scanning electron microscopy, which is applied to the use of wave/particle radiation for material analysis, measuring devices, instruments, etc., can solve problems such as difficulty in achieving satisfactory observation results, test failures, broken inserts, and conductivity of inserts. Achieve the effects of avoiding slipping, deformation or even loss, eliminating potential safety hazards, and reducing discharge phenomena

Pending Publication Date: 2020-11-20
ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Scanning electron microscopy (SEM) is an important means for observing and studying the ultra-microscopic morphology of fungal surfaces. However, the conventional SEM preparation method used to observe fungi in the past, because the fungal sample itself is a monolayer of cells, its main components are fungal spores and hyphae. In the process of sample preparation, it is easily deformed, causing artificial artifacts, and it is difficult to achieve a satisfactory observation effect
In addition, the glass insert used in the insert method is fragile and has poor conductivity, which greatly affects the test progress and effect. A patented method for preparing filamentous fungus environmental scanning electron microscope samples (CN104928343A) succeeded in using edible aluminum foil instead of glass inserts. Solved the problem of broken blades and enhanced the conductivity of the blades
However, the author of the present invention found that the organic acids produced by some fungi during the culture of inserts easily corroded the aluminum foil, resulting in the failure of the test
In addition, the aluminum foil is soft and easy to deform, which is not conducive to the observation of fungi by scanning electron microscopy.

Method used

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  • Scanning electron microscope sample preparation method suitable for fungi
  • Scanning electron microscope sample preparation method suitable for fungi

Examples

Experimental program
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Effect test

Embodiment 1

[0025] 1. Preparation and sterilization of solid medium

[0026] Use deionized water to boil ordinary potato dextrose agar medium (PDA), add 1.5g of agar powder per 100ml, put it into a pressure cooker for sterilization, the sterilization temperature is 121°C, and the sterilization time is 20min. The solid culture is based on the ultra-clean bench and poured into a glass petri dish with a diameter of 9cm, and the thickness of the medium is 4-6mm.

[0027] 2. Preparation and sterilization of stainless steel mesh

[0028] Select 304 stainless steel wire with a diameter of 0.08mm and weave it into a stainless steel mesh with an aperture of 100 mesh, a thickness of about 1mm, and a diameter of about 0.8mm by plain weave method. After being sealed and packaged in newspaper, it is autoclaved at 120°C for 20 minutes, and dried for later use.

[0029] 3. Inoculation and insertion

[0030] Carry out fungal inoculation in the ultra-clean workbench, inoculate the fungus cake (φ=5mm) at...

Embodiment 2

[0041] 1. Preparation of solid medium

[0042] Use deionized water to boil ordinary potato dextrose agar medium (PDA), add 1.5g of agar powder per 100ml, put it in a pressure cooker for sterilization, the sterilization temperature is 121°C, and the sterilization time is 20min, the solid culture after sterilization Based on the ultra-clean bench, pour it into a glass petri dish with a diameter of 9 cm, and the thickness of the medium is 4-6 mm.

[0043]2. Preparation of glass slides and aluminum foil

[0044] Take glass coverslips for tissue observation, cut into 8mm length and width aluminum foil sheets for microwave use and stainless steel mesh sheets, wrap them with filter paper and sterilize at 121°C for 20min, and dry them for later use.

[0045] 3. Inoculation and insertion

[0046] Inoculate the fungus in the ultra-clean workbench, inoculate the fungus cake (φ=5mm) at the center of the medium, insert a sterilized glass slide / aluminum foil on the medium, the distance be...

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Abstract

The invention discloses a scanning electron microscope sample preparation method suitable for fungi. The preparation method comprises the following steps: (1) preparing and sterilizing a stainless steel woven mesh; (2) inoculation and insertion: carrying out fungus inoculation in an ultra-clean workbench, inoculating a fungus cake to the center point of a solid culture medium to be flatly attachedto the surface of the culture medium, and placed into 2-5 sterilized stainless steel woven meshes, wherein the distance between the stainless steel woven meshes and the center point of the culture medium is 3-4 cm; and sealing with a sterile film, culturing in an incubator under proper conditions, and stopping culturing when the exposed stainless steel woven mesh is covered by hyphae; and (4) fixing, dehydrating, replacing and drying the stainless steel woven mesh sample.

Description

technical field [0001] The invention belongs to the technical field, and in particular relates to a sample preparation method suitable for fungal scanning electron microscopy. Background technique [0002] As an important experimental material, fungi are widely used in microbial experiments. Correct culture and observation methods can faithfully reflect their essential characteristics and provide a solid foundation for the classification and identification of fungi. The insert culture method is a basic method for laboratory observation of fungal morphology. The method is simple and easy to understand, and has a wide range of applications. In the microscopic observation of the fungal morphology, it can ensure that the basic morphology of the fungus is not destroyed, and the structure is complete. The basic steps are: pour the solid culture medium into a clean petri dish with a thickness of about 0.3-0.5 mm, and inoculate the inoculation block in the center of the solidified ...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): G01N23/2251G01N23/2202
CPCG01N23/2251G01N23/2202
Inventor李春霞常圣鑫李春泽胡美姣李敏高兆银弓德强母军霞
OwnerENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI