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Quick preparation method for candida albicans specimen for field emission scanning electron microscope

A technology of electron microscopy and Candida albicans, which is applied in the preparation of test samples, material analysis using wave/particle radiation, and measuring devices, etc., can solve the problems of large impact on sample surface structure, cracking, and large damage. Achieve the effect of eliminating surface damage of bacteria and avoiding surface cracking

Inactive Publication Date: 2017-09-01
CHENGDU UNIV
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Problems solved by technology

[0009] The traditional method uses different concentrations of gradient ethanol for dehydration, and the different dehydration time has a great influence on the surface structure of the sample. The observation results show that when the sample is dehydrated in a gradient of 10 minutes per step, the surface of the bacterial biofilm is cracked, or shrunk and sunken to varying degrees. is broken (as in Figure 4 ), and the damage

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  • Quick preparation method for candida albicans specimen for field emission scanning electron microscope
  • Quick preparation method for candida albicans specimen for field emission scanning electron microscope
  • Quick preparation method for candida albicans specimen for field emission scanning electron microscope

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[0029] The specific implementation of the present invention will be described in conjunction with the examples.

[0030] 1. The preparation method of Candida albicans suspension

[0031] Pick out the Candida albicans strains stored on a solid slope in a refrigerator at 4°C, inoculate them on a solid potato dextrose agar (PDA) medium in an ultra-clean workbench, streak and culture them in a constant temperature incubator at 30°C After culturing in medium for 12 hours, pick a single colony on the petri dish and put it in a liquid common broth medium, and culture it in an air bath shaker at 30°C at 200rpm for 12-24 hours to obtain a suspension of Candida albicans;

[0032] Then centrifuge at 8000rpm for 3-5min, discard the supernatant, and dilute the cell pellet to 10% with sterilized liquid potato dextrose agar (PDA) medium. 5 ~10 6 cfu / mL was used as the standard test bacterial suspension, and stored in a refrigerator at 4°C for later use.

[0033] 2. Preparation method of C...

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Abstract

The invention belongs to the bio-technical field, and particularly discloses a quick preparation method for a candida albicans specimen for field emission scanning electron microscope. The quick preparation method is characterized in that candida albicans thalluses are added into a glutaraldehyde fixing solution for being fixed; sterile deionized water is adopted to clean; and tertiary butanol with different concentration gradients is adopted for dehydration; a bacteria solution is dropped into pure tertiary butanol to freeze in a refrigerator of 4 DEG C; and after a dried specimen is sprayed with gold in an ion sputtering mode, the morphological structure of a thallus biological membrane can be observed through a scanning electron microscope by regulating and setting parameters. According to the quick preparation method for the candida albicans specimen for field emission scanning electron microscope provided by the invention, excreta of the candida albicans is reduced to affect image-forming quality of the specimen; a phenomenon that some samples are covered and shielded by phosphate particles is eliminated to the greatest extent; tiny deformation of the specimen is reduced and the biological membrane is prevented from dropping slightly; the surface of the thallus biological membrane is prevented from cracks or shrinkage sinking of varying degrees and even breakage; and meanwhile, damages on the surfaces of the thualuses are eliminated.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a rapid preparation method for Candida albicans field emission scanning electron microscope samples. Background technique [0002] The biofilm of Candida albicans is also called biofilm. In nature, the vast majority of bacteria exist in biofilms. Studies have shown that the existence of some Candida albicans biofilms is related to the severity of their pathogenesis. Therefore, exploring the biofilm structure of Candida albicans has gradually become a focus of scientific researchers. [0003] Scanning electron microscopy has become a powerful tool for studying the microscopic world, and many major discoveries and research results are obtained through observation. However, in the actual observation operation of the instrument, because the scanning electron microscope is a high-precision instrument, the structure is complex, and there are many factors that affect its imaging. In orde...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N23/225G01N1/28
CPCG01N23/2251G01N23/2202
Inventor 宋明珠茆灿泉李建苏杨黄婵黎浩峰周希陈渝姣蒲琪彭正伟雷子叶
Owner CHENGDU UNIV
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