[Histonet] Has anyone done a comprehensive study on fixatives?

Rittman, Barry R Barry.R.Rittman <@t> uth.tmc.edu
Sat Jun 24 07:52:26 CDT 2006

I think that the problem with comparing fixatives is that  there are so many thousands of formulae and processes in the literature that it is impossible for the newcomer to know where to start. Everyone has their favorite technique but often with their mown varaitions to the original.
The following all play a role in the final image:
1.  An individual fixative will give variable results with different tissues and tissue components.
2. Most laboartories will have different sizes and volumes of tissues.
3. Different processing techniques and times are used.
4. There is the question of secondary fixation, if not deliberate at least during the subsequent processing.
5.If wax processing then there maty be different reagents used for removing wax etc.
6.Different images will result sometimes with even slight variations in the staining, histochemial or immunohistochemical techniques.
7.Not all reagent do what the supplies states, and this appears at least to me to be especially true with immunohistochemistry reagents.
8. Sometimes histotechs, for a variety of reasons do not use adequte controls for specificity of reactions.
9. A positive or negative result may be in the eye of the beholder whether histotech or pathologist.
10. Many individuals who write to Histonet are using tissues from different species and it may not be possible to use the same technique for all species, whether  this is regarding fixation or processing or staining.
I feel that the most important criteria are conisitency and controls.
Any publications regarding fixation should include  not only the results after fixationn but also after procesing.
If you want to compare fixatives you are best to use publications as a starting point and thedo a trial run yourself with the tissues you want to use.
First decide precisely what you wish to finally demonstrate. 
You should include some different times and keep complete notes about temperature, concentrations etc. at all steps including, fixation, processing etc.


From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Lee & Peggy Wenk
Sent: Sat 6/24/2006 1:35 AM
To: 'Cote, Laurie'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Has anyone done a comprehensive study on fixatives?

I don't have the issue in front of me, so maybe a Histonetter can supply
more information.

A couple of years ago, the NSH Journal of Histotechnology published a
special September issue, with all the articles related to fixation. There
were some good charts in it, as to what type of fixative fixes what

Contact NSH 301-262-6221, and see if they have any old issues you could buy.
(The Journal archive web pages are still under construction on the NSH web

Peggy A. Wenk, HTL(ASCP)SLS
William Beaumont Hospital
Royal Oak, MI 48073

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Cote, Laurie
Sent: Friday, June 23, 2006 9:09 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Has anyone done a comprehensive study on fixatives?

Has there ever been a comprehensive study done on fixatives?  For instance,
the differences between fixation with formalin, different kinds of alcohol
or other fixatives? Specifically, is one fixative better for IHC?  If a
paper has been done and published, anyone know where it might be?  I tried a
pubmed search and was relatively unsuccessful.

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