I have been using the following protocol for preparing mouse kidneys for cryosectioning:
1. Perfusion with 4% PFA
2. Fixation O/N in 4% PFA
3. 30% sucrose O/N
4. 30% sucrose O/N
5. Embedding in OCT
6. 5 uM sections
My H&E stains show spaces between structures in the tissue. Could there be something wrong with the protocol I am using to prep the kidney before embedding? I did read that bringing the kidney into 30% sucrose immediately might impact tissue morphology. If anyone could shed some insight into the proper protocol I would be grateful!
Probably go for Paraffin embedded blocks. For large tissue, penetration of sugar could be an issue.
Paraffin embedded sections are more stable than the cryosections. for the preparation of the H&E I prefer to use Paraffin sections. It will give great results.
The cryosections can be better for the detection of specific antigens. That cant be detected by the Paraffin sections...
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