My question pertains to human heart tissue previously embedded and frozen in OCT.
We have recently cut sections of varying thickness (100-1000 um) for processing and staining while free-floating. The sections were cut on a cryostat and transferred immediately to formalin for fixation. However, while the thicker sections remained intact, the thinnest section, ~100 um thick, seemingly completely disappeared following 2 hours of fixation. Has anyone previously observed the dissolution of very thin tissues in formalin?
Thanks.
No, the sections can not disappear. Section will not dissolve in 10%NBF. You can keep as much as you want in 10%NBF.
First of all, you should ask a pathologist if cryosectioning is appropriate for your staining. Paraffin embedding is usually the good choice for routine histological (e.g. H&E) stainings.
Then, if cryosectoning is the adequate method for the staining, your tissue samples should be fixed in formalin and dehydrated in sucrose before cryosectioning.
For detailed protocol, you can check our recent study
https://akademiai.com/doi/abs/10.1556/004.2018.026
Formalin will not dissolve tissue. Most likely, your thinner sections only included OCT and no tissue, or maybe they were lost somewhere in the process, perhaps because their thinness made them less visible.
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