I need to take samples of porcine longissimus dorsi and semimembranosus muscles for immunohistochemistry analysis. However I can't find any detaild protocol or explanations even in the relevant papers.
Will these be postmortem muscle samples or are these percutaneous biopsies from living pigs? Do the primary antibodies work on fixed tissue or only on unfixed tissue?
Gordon L Warren This will be postmortem sampling. I need a detailed procedure/instructions for the slaughterhouse workers how to cut the muscle from the carcass, like which part of latissimus dorsi and semimembranosus to take, which tool to use.
I am going to freeze the tissue and store on -80 and use it to prepare cryosections and to isolate RNA ( I have this protocol).
Also, I would really appreciate if you can give me advice regarding the sample transport since I will not be able to freeze it onsite, however I need to ensure high quality cross-sections for IHH.
Will you not be present for the tissue processing? Freezing muscle for cutting cross-sections in a cryostat is not that easy, at least not for a beginner. For one, the tissue sample should be relatively small, i.e.,
Gordon L Warren Thank you very much for your detailed answer. However, I already have the protocol for freezing the samples properly (experience from chicken and human samples), my question was directed to dissection part, how to recognize semimembranosus in pig and what is proper way to take peace of tissue, how big, which tool to use (knife, razor blade...). Of course first cuts will be cut in smaller adequate pieces before freezing.
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