Pre fixation is first done by Glutaraldehyde, post fixation with osmium tetra oxide is then required and finally dehydration with ethanol. After that specimen is glued to SEM stub by carbon tape, finally dried in vacuum, and sputtered by carbon or gold or platinum. Below are some varied techniques in literature very briefly:

(1) The Whatman strip containing the fungus was fixed with glutaraldehyde (2.5%, v/v) in phosphate buffer (10 mM; pH 7.4) for 4 h. The fixed samples were rinsed twice with deionized water and the samples were dehydrated with increasing concentrations of ethanol (10, 20, 40, 60, 80 and 100%) for 10 min each. After dehydration, the samples were dried at room temperature for 3 h. The dried samples were sputter coated and visualized as described above.

(2) The fungal sample was first fixed with glutaraldehyde in phosphate buffer (10 mM; pH 7.4) for 30 min followed by washing with doubled distilled water four times and treated with osmium tetroxide (0.1% w/v) for 30 min. The samples were dried at room temperature for 3 h. The dried samples were sputter coated and visualized as described above.

Reference:

https://dx.doi.org/10.1038%2Fs41598-018-34629-8

Pre fixation is first done by Glutaraldehyde, post fixation with osmium tetra oxide is then required and finally dehydration with ethanol. After that specimen is glued to SEM stub by carbon tape, finally dried in vacuum, and sputtered by carbon or gold or platinum. Below are some varied techniques in literature very briefly:

(1) The Whatman strip containing the fungus was fixed with glutaraldehyde (2.5%, v/v) in phosphate buffer (10 mM; pH 7.4) for 4 h. The fixed samples were rinsed twice with deionized water and the samples were dehydrated with increasing concentrations of ethanol (10, 20, 40, 60, 80 and 100%) for 10 min each. After dehydration, the samples were dried at room temperature for 3 h. The dried samples were sputter coated and visualized as described above.

(2) The fungal sample was first fixed with glutaraldehyde in phosphate buffer (10 mM; pH 7.4) for 30 min followed by washing with doubled distilled water four times and treated with osmium tetroxide (0.1% w/v) for 30 min. The samples were dried at room temperature for 3 h. The dried samples were sputter coated and visualized as described above.

Reference:

https://dx.doi.org/10.1038%2Fs41598-018-34629-8

Apply a small volume of the suspension of bacteria or fungi on to millicell(millipore) filter of pore size 0.22 micron or 0.4 micron. You can also use nuclepore membrane. Let it stand for sometime or give gentle spin in a centrifuge to get the buffer solution out. Rinse, dehydrate, dry in HMDS(NOT vacuum dry) and do the rest as above. Check SEM references.

https://ar.ovalengineering.com/scanning-electron-microscope-microbiology-138749

English translation please.

Thanks.

Preparation of Microfungi for Scanning Electron Microscopy

link.springer.com › content › pdfThe scanning electron microscope (SEM) has broad applications to fungal re- ... generated by sample preparation procedures, since none are artifact-free, and ... ration of bacteria (Watson et aZ., 1980) and conidial fungi (Cole, 1981b). The.

Firstly, you must prepare a very small amount of the suspension of the microorganism which you want to test it such as fungi or bacteria to millipore filter with size of 0.22 micron or less.

Dear Yeshaneh Adimasu you can find a few more answers to your question when you check out the following closely related RG thread entitled "How to prepare fungal specimens for Scanning Electron Microscope?":

https://www.researchgate.net/post/How-to-prepare-fungal-specimens-for-Scanning-Electron-Microscope

Please also see the attached papers entitled "Scanning Electron Microscopy (SEM) Protocols for Problematic Plant, Oomycete, and Fungal Samples" and "The scanning electron microscope in microbiology and diagnosis of infectious disease"