[Histonet] Frozen section artefact

Hobbs, Carl 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.

Curious always,


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