[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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