[Histonet] Fixed frozen non-paraffin mouse brain

[Porter, Amy]

You need to add sucrose to your PFA we use 4%PFA+4%Sucrose to fix and then cryoprotect in 30% sucrose which all need to be prepared in Phosphate buffer solution NO saline .... if you message me directly I would be happy to share our SOP this was a very hard thing to learn.... not a lot in technical references about this process.

Sent from my iPhone

> On Jul 4, 2020, at 9:09 PM, Roy, Edward J via Histonet <histonet at lists.utsouthwestern.edu> wrote:
> 
> As a research lab, we sometimes would like to use paraformaldehyde-fixed but non-paraffin embedded tissues; paraffin embedding alters antigens and necessitates antigen retrieval, but simple fixation does not. We have done the traditional 30% sucrose before OCT and freezing, with cryostat sectioning, but results are inconsistent, sometimes producing Swiss-cheese brains. Does anybody have an alternative to 30% sucrose that is more reliable?  I didn’t see anything in the Archives after a search for “30% sucrose”.
> Thanks very much,
> Ed Roy
> 
> 
> Edward J. Roy, PhD
> Professor Emeritus
> Department of Molecular and Integrative Physiology
> University of Illinois at Urbana-Champaign
> Urbana, IL 61801
> 217 333-3375
> 
> 
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