[Histonet] Frozen section artefact

[Adam Boanas]

Thanks for the reply - Previously I was going directly from the fixative to the PBS. Yesterday I tried air drying the sections for 30mins prior to PBS and the sections looked much better. Will try increasing the alcohol content as you suggest.
Adam

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Hobbs, Carl
Sent: 25 September 2012 18:37
To: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Frozen section artefact

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,



Carl






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