[Histonet] Intestinal Tissue Questions

[Favara, Cynthia (NIH/NIAID)]

Wendy,

You have gotten fabulous advice. I only add what I do. I perfuse the mouse
as we are studying brain and this is far superior. I leave the gut intact in
the animal. Cut just beneath the stomach, and at the cecal junction, insert
a dulled needle per Gayle Callis rinse jelly roll and put on lens paper. Do
the same for the lower intestines. Can do all in one but I find this easier.
Get some normal mice try everything and practice, practice, practice..pretty
soon you will be giving advice.

Cynthia Favara
NIAID/NIH/RML/LPVD
903 South 4th Street
Hamilton, MT 59840
406-363-9317

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-----Original Message-----
From: wbedale <@t> biochem.wisc.edu [mailto:wbedale <@t> biochem.wisc.edu] 
Sent: Friday, January 21, 2005 9:32 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Intestinal Tissue Questions

Hi to all of you,

I'm quite new to histology and have a few basic questions.

Here's what I'm doing:

Tissue:  mouse intestine;  small and large intestine is harvested, cut open
longitudinally, and rinsed with PBS prior to fixation; 3 cm lengths are
fixed.
Fixation: 10% formalin
Processing and embedding:  manual processing, paraffin embedding, 4-6 micron
sections
Staining:  H&E staining

Two questions:

1.  After fixation, is it better to store my tissue (for ~1 month or so) in
fixative, or in 70% ethanol?

2.  I'm attempting to get transverse sections of the intestine;  sometimes I
notice that my tissue ends up inside out (mucosa facing outward).  I think
it may happen when I use a razor blade to cut the tissue longitudinally vs.
using scissors to make the longitudinal incision, and am going to test that.
Has anyone had any experience with this?

Thanks for any advice.

Wendy

Wendy Bedale, Ph.D.
Assistant Scientist
Department of Biochemistry
University of Wisconsin-Madison
433 Babcock Dr.
Madison, WI 53706
608-262-3099 ext. 3266
wbedale <@t> biochem.wisc.edu


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