[Histonet] fixation question for IHC

[Anatoli Gleiberman]

Jan,
Depends on your material, further processing and antigens. Surprisingly, some antigens are better preserved after formaldehyde fixation than after buffered formalin (probably, due to high sensitivity of particular epitope to alcohol traces from standard formalin solution). I know at least one - mouse EpCAM epithelial marker almost negative after formalin and quite well stained after formaldehyde (rat monoclonal anti EpCAM antibody G8.8 from Developmental Studies Hybridoma Bank). Morphology and antigen staining on cryo-sections from formaldehyde-fixed mouse embryos (e11-e16) are slightly better than after formalin fixation (used the same 4-6h fixation time either with buffered formalin or with 4% formaldehyde on PBS prepared from 20% formaldehyde water solution from Electron Microscopy Sciences, following PBS wash and storage in PBS at +4 C). It seems, embryonic tissue is quite sensitive to coagulation induced by the presence of small amount of alcohol. 

Anatoli Gleiberman, PhD
Director of Histopathology
Cleveland Biolabs, Inc
73 High Street
Buffalo, NY 14203
phone:716-849-6810 ext.354
fax:716-849-6817
e-mail: AGleiberman <@t> cbiolabs.com

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jan Berry
Sent: Friday, January 28, 2011 1:15 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] fixation question for IHC

Is there a difference between using paraformaldehyde and neutral buffered formalin when choosing a fixative for IHC?  I would prefer to use formalin because of easier preparation, but am willing to put in the extra time to make fresh paraformaldehyde solution if there is a compelling reason.

Jan Berry
University of Michigan
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

 


This communication may contain privileged information.  It is intended solely for the use of the addressee.  If you are not the intended recipient, you are strictly prohibited from disclosing, copying, distributing or using any of this information.  If you received this communication in error, please contact the sender immediately and destroy the material in its entirety, whether electronic or hard copy.  This communication may contain nonpublic information about individuals and businesses subject to the restrictions of the Gramm-Leach-Bliley Act.  You may not directly or indirectly reuse or redisclose such information for any purpose other than to provide the services for which you are receiving the information.