[Histonet] Rodent processing and artifactual spaces

James Watson JWatson <@t> gnf.org
Fri Aug 3 11:16:44 CDT 2012

Something that I have done for years on animal tissues is: instead of 100% ETOH on the processor we use 5% glycerin in 100% ETOH.  This keeps the tissue from becoming brittle and greatly decreases soaking time while cutting.  Started doing this in the 80s at AFIP when processing 5 cm thick whole dog heart cross sections that took 48 hours to process.  2 minor problems with this:  1) does not work on fat, so we keep extra containers of 100% ETOH to switch to when processing fat. 2) we have to rinse our xylene recycling container and recycler monthly with 100% ETOH.

James Watson HT  ASCP
GNF  Genomics Institute of the Novartis Research Foundation Tel    858-332-4647 Fax   858-812-1915 jwatson <@t> gnf.org

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Grantham, Andrea L - (algranth)
Sent: Thursday, August 02, 2012 4:43 PM
Subject: Re: [Histonet] Rodent processing and artifactual spaces

I process rodent tissue among other types of tissues and the times are similar to those that I use. In the place of Xylene I use Clear Rite 3. I find that this is a little more gentle and the tissue while still dry is not as bad and some things I don't even have to soak - like ovaries and some brains. But when I do soak I use a little glycerin in my water and let the faced blocks sit on the cold plate for 30 min-1 hr. Usually that is all it takes to get really pretty sections.
One other thing to think of is the temp of your waterbath. I keep mine around 40 degrees C.
I let the slides dry completely before putting them in the oven before staining. If you have water under the sections it could produce the some of the spaces that you are seeing when the water expands in the oven.

Andrea Grantham, HT (ASCP)
Senior Research Specialist
University of Arizona
Cellular and Molecular Medicine
Histology Service Laboratory
P.O.Box 245044
Tucson, AZ 85724

algranth <@t> email.arizona.edu<mailto:algranth <@t> email.arizona.edu>
Tel: 520.626.4415     Fax: 520.626.2097

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