Hi guys,
I did an Epon embedding of mouse liver for trasmission electron microscopy, in order to see mitochondria cristae, but the images appear blurry and I cannot see the cristae properly (see the attached image). It is very strange, because usually I get very good images from other tissues. I fixed the tissue in GA 2.5%+PFA 2% in 0.1M sodium cacodylate ON at 4 degrees, then post fixed in OsO4 1% 2hr at 4 degrees and then embedded in EPON
I will really appreciate if somone could give some advices, to overcome this problem.
Should I add something like tannic acid, or potassium forrocyanide, or something else?
Many thanks
Francesco
Oh how we all love the occasional inconsistencies of TEM processing...
In general I do not like the results with tannic or gallic acid too much, whether applied before or after osmication. I prefer to work with reduced osmium (using buffered 1.5% Ferricyanide +1% OsO4), which results in better membrane staining.
You can also try the good old OTO method, which is a bit more time consuming, but will definitely help with increasing membrane contrast.
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