I'm trying to do histology on whole rat knees and am wondering if anyone had any idea as to how long it needed to be left for each change in Histo-Clear, or if it can be left overnight? When doing it previously the samples have become very brittle and hard to section and I'm thinking maybe the Histo-Clear is over hardening them?
How about looking in the literature to find author(s) who have done such same?
Minimum time, 3 changes, 3 minutes each,
I have left the slides in Histoclear II for overnight, there is no problem, I hope that answered your questions.
Good luck
If you are processing them for paraffin histology, you have two changes in histoclear, at most 1 hour each. You declare victory when your specimens are transparent (in a yellowish kind of way). Any longer than that at they will harden. I also suggest intermediate steps of ETOH/Histoclear before, and Histoclear/paraffin afterwards. The only time I would ever leave anything overnight is in alcohols, 100% ETOH, and even then, in the refrigerator. If you are staining, then it doesn't matter how long the slides are in histoclear.
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