[Histonet] Re: Histonet Digest, Vol 108, Issue 9

[Madeleine Huey]

> Message: 4
> Date: Tue, 6 Nov 2012 11:41:26 -0800 (PST)
> From: Amy Lee <amylee779 <@t> yahoo.com>
> Subject: [Histonet] How to fix frozen section?
> To: histonet <histonet <@t> lists.utsouthwestern.edu>
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> Hello,
>
> I will receive some frozen tissue sections that are not fixed. I rarely do IHC on frozen sections. I searched some articles about fixation. Some use 50/50 of acetone/alcohol and some use only alcohol. Could you please recommend a fixation method for me?
>
>
> Thanks,
>
> Amy
>
Amy,

Here's my method (based only on personal experiences);

1st) Remove frozen tissue section/s from freezer
2nd) Immediatly pre-fix in 10% NFB @ RT for 10 min (do NOT thawed
slides @ RT) "I found some protein leak out if pre-fix with Acetone
without 10%NFB"
3rd) Re-fix in Acetone for addition 5min - 10min @ -20c
4th) Air dry slides with fan for ~ 5 min (or RT for 10 min).  "I
purchase $10 portable fan from drug store"
5th) Wash off any excess OCT with washing buffer 3x
6th) Proceed your IHC staining......

Good Luck!
Madeleine Huey BS, HTL (ASCP) QIHC
Supervisor - Pathology
madeleine_h <@t> elcaminohospital.org