I am performing histology on rat knee joints and would like to look at the changes in cartilage, I have fixed the joints in 10% NBF for 3 days and decalcified for 11 days in 10% formic acid. What is the best time intervals for complete dehydration of the sample using ethanol?
Thank you.
When I did mouse knees I left them for an hour for every dehydration step. If you are in no rush you can start in the morning with 30% EtOH and after an hour and a half, when you have time, you change it to 50%, then 75%, then 95%, then 100% and again 100% and leave them in 100% overnight. Use excess volume for each step, a 50 ml falcon tube for each rat knee work well and makes it easy to work with.
I would also suggest that you occasionally gently agitate/flip the tubes. Mixing of fluids is useful in large samples. Diffusion alone is too slow.
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