I am concerned that formalin fixed paraffin embedded mouse eyes with the lens and cornea in tact, will present sectioning problems, as well as shrinkage causing the retina to detach. I found a whole eye protocol using phenol in the lower alcohols (70% and the first 95%) that claims to soften the sclera and lens, and using chloroform instead of xylene as the clearing agent to reduce shrinkage. Does anyone have experience with this? I think the protocol is for human eye. Should I reduce the times in each reagent for the much smaller mouse eye, or would times not matter? I would be processing with this schedule manually, since I don't want to introduce the chloroform or phenol into my automatic tissue processor.
Also, I have some eye cups (with lens and cornea removed) that have, in the past, shown detachment of the retina upon sectioning of paraffin blocks. These were fixed with 10% formalin and processed routinely. Should I switch to Bouin's to reduce shrinkage? or Davidson's (does this reduce shrinkage?).
Thanks so much,