I am requesting your kind assistance in proper steps in harvesting, decalcifying and embedding the knee joint samples, assuming that sectioning and histological staining of the knee samples are similar with other kinds of tissue. Thank you.
For LM use 10% tetrasodiumEDTA to decalcify after fixation with Bouin's fluid. Embed in paraffin. For EM use my protocol in the paper by Warshawsky and Moore, that is enclosed. HW
Dear Kok Yong,
it would be helpful too if you could give some detail about your (anatomical=dissection) preparation technique and the objective of your study/studies.
The most important point (I guess) is that you must have fixed totally your specimen to be able to decalcify your whole specimen by EDTA, or incubation in suited acids (to avoid maceration of the tissue).
The (primary) fixative/fixation will / can depend on the task you are to reach / thinking of... instead of Bouin's fluid (which nowadays seems to be a bit critical due to the use of "picric acid" very often said to be dangerous because of the possibility to explode [NOTE: only when the chemical is drying out] so you certainly know that you can use also NBF (neutral buffered formalin=formaldehyde=FA) as a primary fixative . Or - if you intend perhaps also to use part of the specimens afterwards for electron microscopical purposes / ultrastructural examination you can penetrate your specs in a mixture of buffered FA-GA=glutardialdehyde-fixative (e.g. 1-4% FA, 1-5% GA in 0.1M - 0.13 M [Millonig's] phosphate buffer.
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