> Message: 4 > Date: Tue, 6 Nov 2012 11:41:26 -0800 (PST) > From: Amy Lee <amylee...@yahoo.com> > Subject: [Histonet] How to fix frozen section? > To: histonet <histonet@lists.utsouthwestern.edu> > Message-ID: > <1352230886.68993.yahoomailclas...@web126002.mail.ne1.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > 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 madelein...@elcaminohospital.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
