Hi
Is there a product that I can use to visualize microorganisms sitting on a solid 5x5 mm white surface without electron microscopy? I have tried EM without luck because it only shows a small bit of the structure and its dificult to distinguish between the surface and the microorganisms (mostly bacteria). I would like to see how the microorganism is distributed on the surface . Does anyone have experience with this? Is there a staining technique outh there that I can use?
Well, if the solid surface is thin enough to be able to use an epifluorescence microscope to visualise it (i.e. like a microscope slide), then you should be able to use a DAPI staining protocol or Sybr green protocol to stain them.
You should use the surface as a slide and mount it with a cover glass, so a way to prevent your bacteria to move is by adding some agarose (I think 0.1-0.05% don't remember well now).
If it is hard to proceed with any protocols since that microbe may be washed-out. You can try tetrazolium as an indicator of viable cells and confirm under the fluorescence microscope. No washing steps necessary. But only viable cells can be detected.
It's on a stone surface so I'm looking for a method that only utilizes stereomicroscope. Could an alternative method be putting the stone into a resazurin solution and measure viability?
Ultra-microscopy (dark-field microscopy) can show unstained particles the size of bacteria, but will not resolve their structure, i.e., it will be difficult to tell whether these are bacteria or just dust. See for example doi:10.1038/srep20729.
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