RE: Quality of Fixation

From:"Morken, Tim" (by way of Histonet)

Charles, the use of a vimentin antibody (specifically clone V9) has been
used to determine the quality of formalin fixation for immunohistochemistry
(Battifora H., Am J Clin Pathol 1991; 96:669-671). It is possible you could
use the same method to show that your fixation method is "at least" as good
at other methods. Briefly, since vimentin is ubiquitous, and is suseptible
to over-fixation by formain (reduced staining if over-fixed), then staining
for vimentin will show whether proper fixation has occurred.

Good luck!

Tim Morken
CDC, Atlanta

-----Original Message-----
From: Charles W. Scouten, Ph.D. []
Sent: Thursday, March 06, 2003 10:27 AM
Subject: Quality of Fixation

I need some help from the experts.

Is there a standard measure of quality of fixation?  A stain that shows
degree of fixation?  I don't have a lot of equipment, and don't want to
spend a lot of money, but I have been challenged by a reviewer to show that
my perfusion methods (animal research) results in as good a fixation as
standard methods.

I can see it does, tissue quality is excellent for sectioning and staining,
but how do I quantify or show that?  Age it 90 days and report degree of
crumbling?  It is 4% formaldehyde fixation.


Charles W.  Scouten, Ph.D.
5918 Evergreen Blvd.
St. Louis, MO 63134
Ph: 314 522 0300
FAX  314 522 0377

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