Methods and Techniques Discussion > General Histology

Paraffin embedded Whole mouse eyes, eye cups

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pweston:
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,

Paula

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