[Histonet] Will brain tissue shrink even more if stored in 70% alcohol prior to processing?

CM Bush clarissabush <@t> sbcglobal.net
Thu May 26 11:39:54 CDT 2005


Dear Histonet,
 
Hello, here is my first post to the list, thank you in advance for your help.
 
Summary:
Having problems with mouse and human brain tissue shrinkage durning processing, but would like to better preserve antigens, concerned storage of tissue 70% ethanol will shrink tissue even further: 
 
We have lots of brain specimens, stored in formalin for a long time (months up to 10 years), both human and mouse.  We perform immunohistochemistry on paraffin embedded brain.  Of course, there is the massive over fixation. Also the tissue shrinks a lot during processing.  
 
In my last lab, after brain tissue had been fixed in formalin for 7 days we would store the tissue in 70% ethanol.  I'd like to start storing the brain tissue I work with now in 70%, but I'm worried about the tissue possibly shinking too much, as the tissue already seems to shrink by greater than 60% after processing.
 
(Previously, we would gradually increase the alcohols, 30% for 1 hour, then 50% for an hour in a bucket, room temperature on a rocker table, then put the cassettes into a bucket of 70% and store at room temp or 4C- does this process help any with controling the shrinkage factor?)
 
Maybe this is a little bit long...thank you very much for your time.
 
CM Bush
 
 
 




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