I hope to make some frozen sections from adipose tissue. However, my adipose tissue has been stored in liquid nitrogen for months. Is that OK? If yes, what are the notes?
In order to prevent loose of morphology, I would 'warm' it slowly (-80°C, then -20°C).
Dear Ting, is that human tissue? What species are the antibodies generated in?
Dear Ting Yu,
I have had to do this for my current study using mouse tissue. I gradually defrosted the samples and then embedded them in OCT.
I have used primary antibodies from goat and rabbit and they have worked really well.
Goodluck!
My experience was with formalin/paraffin. I defrosted the tissue, then fixed it for 48h in 4% buffered formalin, followed by routine histological dehydration, xylene and embedding in paraffin. I have never done frozen cuts like Brooke, but I agree with her. Epitopes are preserved in -80°C, thus you shouldn't have problems with IHQ or IF procedures.
I work with C57Bl/6 mice samples that are usually pretty small, unless they have ate a obesogenic diet. My routine is 60 min/bath, in the following order: tap water 5 min to remove excessive formalin (usefull when you plan to do IHQ), then ethanol 70% x1, 80% x1, 95% x1, 100% x3, xylene x2, paraffin x2. It tooks about 11h (if you consider the embedding time!).
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