Re: hair cell IHC

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From:Tony Henwood <>, (Karen Larison)
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Dear Karen,

> One of the post docs here is doing IHC on frozen sections of frog sacculi.  The 
> staining works well, but the morphology of the hair cells is horrible.  Maybe someone 
> has a suggestion on how to preserve the integrity of these cells though the 
> cryosectioning/staining process.  We embed the sacculus in agar, sink in 30% sucrose, 
> and freeze slowly over liquid nitrogen.  For most small specimens we work with, this 
> procedure works well, but the sacculus seems to be more picky-you-nish.  So if anyone 
> out there works with this organ, and has ideas on how to improve our morphology, I'd 
> love to hear from you.

Maybe the poor morphology is due to the post staining dehydration, 
clearing and coverslipping. Try coverlipping straight from water 
using an aqueous mountant like Aquamount. It might help.

Regards, Tony
Tony Henwood
Senior Scientist
Anatomical Pathology
Royal Prince Alfred Hospital

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