is there any suggestion to avoid retinal damage during HE staining?
It's very difficult to avoid retinal separation. To start with, the process of dissecting the eyes out probably loosens retinal attachment. Variable shrinking of the different eye components with fixation might play a role as well, as does slicing of the eyeball post-fixation. You don't mention which species you are working on; if it's a species larger than rodents, you can try injecting fixative into the eyeball to speed up fixation, but avoiding putting too much pressure during injection. You can also try head or whole-body perfusion with fixative, which would avoid dissection-associated damage. But in general, pathologists are used to evaluating retinas whether they are detached or not, and there are ways to differentiate post-mortem detachment from pathologic detachment.
well, thank you very much. my work is on mice retina. prolonged fixation may affect the tissue so it can easily be detached?
second, the outlook of the whole retina is not very good. what could be possibilities?
Ethanol will definitely harden the tissue and make it more brittle, if you leave it in for a while. I would keep the post-fixation ethanol period as short as possible, or use formalin instead of ethanol, as this won't cause as much tissue hardening.
I have been doing eye histology for 40 years with no retinal detachment in paraffin sections. 4% PFA, Bouin's, or any aqueous fixative should never be used. The osmolality difference is what causes the retinal photoreceptors to detach from the RPE.
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