I don't know what your heating method is but you might want to try a gentler one than you're using.  Try a steamer or pressure cooker or if you don't have those, try bringing your slides and buffer just up to a simmer in a microwave, then transfer them to a 93-95 degree water bath.  The buffer won't boil this way and you may have better luck with tissue adherence.

Linda A. Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
A4/204-2472
600 Highland Ave.
Madison, WI 53792
(608)265-6596
FAX: (608)262-7174


-----Original Message-----
From: Fernando Capela e Silva [mailto:fcs@uevora.pt]
Sent: Thursday, April 10, 2003 2:45 AM
To: HISTONET@pathology.swmed.edu
Subject: HIER


I use Citrate buffer (PH 6.0) at 95-98#186# for 10 min as HIER. However this
method cause severe damage in some sections and other are completly lost. In
a paper the authors used 14% EDTA at 65#186# C overnight.

I would like if someone help me about this problem and if I is a good choice
using citrate incubation overnight .

With compliments.


* * * * * * * * * * * * * * * * * * * * * * * * *
Fernando Capela e Silva
Laborat#243#rio de Biologia da Conserva#231##227#o
Departamento de Biologia
Universidade de ...vora
Apartado 94
7002-554 ...vora
PORTUGAL

Phone: +351-266 760 800
Fax: +351-266 711 231
Email: fcs@uevora.pt

http://evunix.uevora.pt/~fcs/FernandoCapelaSilva.htm