I have to detect cells in scaffold on SEM. Which method of contrast can i use?
Hi Egor, it depends on what kind of imaging you want to perform.
For secondary electron conventional SEM imaging after you dehydrate the sample (by chemical dehydration or critical point drying) you deposit a thin layer (5-10 nm) of gold by sputter coating (seeArticle Delivery of Brain-Derived Neurotrophic Factor by 3D Biocompa...
.) But if you want to image inside the cells by FIB-SEM than you need to prepare the sample as you would do for TEM imaging and if you want to do SBEM than you should follow a protocol with extra-contrast (seeArticle High contrast en bloc staining of neuronal tissue for field ...
)
Dehydrate your samples and sputter coat them with Au/Pd for 60 seconds.
If your scaffold does not absorb OsO4, you may want to try it. No other contrasting methods that I am aware about. Good news - at right magnifications cells are easy to recognize thanks to their morphology.
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