[Histonet] Frozen section artefact
carl.hobbs <@t> kcl.ac.uk
Tue Sep 25 12:37:01 CDT 2012
Assuming that you are fixing fresh-frozen tissue sections:
Tissue is autolysing.
Use Formalin for 10 mins or 1:1 acetone:alcohol for 10 mins.
Imho, acetone is not a fixative..it's a delipidizer.
5 mins in that mixture is too short a time for the alcohol to be an effective coagulant fixer.
However, a longer time in alcohol may well mask your Ag sites.
I see this artefact in acetone fixed frozen sections, often.
When immunostaining for MHCs in muscle, no fixation is required but, if DAPI/Hoechst is included....nuclear streaming will be seen.
Always try several fixing fluids to get the best results on any new Ab.
I may be wrong....happy to be corrected.
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