RE: [Histonet] Problem with intestinal epithelium

From:"Cervantes, Jessica"

Rene, Patsy, Jennifer, and Ruth- 
I followed this thread with some interest as we have a project in which we'd like to preserve the mucosal layer in rat intestines.  Would any of you be open to sending me a protocol of the steps following the formalin fix?  

I'd like to know specifically what kind of formalin you are using (percentage, buffer), and what stains (if any).  We have tried PAS/Alcian blue staining for the mucous, but since the mucosal layer was destroyed during our sample prep, we only saw mucous in the goblet cells.

Any details you can provide will be much appreciated.
Thank you,
Jessica Cervantes
Bend Research, Inc
Bend, OR 97701
(541) 382-4100

-----Original Message-----
[] On Behalf Of Jennifer
Sent: Wednesday, March 21, 2007 10:10 AM
To: Mikael Niku
Subject: Re: [Histonet] Problem with intestinal epithelium

We get bovine intestine from a local university that has an agriculture 
program.  We provide them with a large container of formalin and ask that 
they open the intestine and place it immediately in the formalin.  We have 
had great preservation of the epithelium.  The piece of intestine is about 
8-10 inches long.

Jennifer MacDonald
Director, Histotechnician Training Program
Mt. San Antonio College
1100 N. Grand Ave.
Walnut, CA 91789
(909) 594-5611 ext. 4884

Mikael Niku  
Sent by:
03/21/2007 03:49 AM


[Histonet] Problem with intestinal epithelium

Dear Histonetters,

we are trying to make good-quality sections (paraffin and cryo) of 
bovine intestines.
The problem is that the epithelium seems to be stripped off whatever we 
do. This seems to be a problem especially with slaughterhouse material, 
although the delay from death to sampling is less than 30 min.

Currently we are cutting the intestine to pieces, washing with PBS 
buffer, and fixing immediately in buffered paraformaldehyde or freezing 
in isopentane / liquid nitrogen.

Any ideas, anyone? Or are we just too late here, could the mucosa be 
destroyed so soon after death?

With best regards,
Mikael Niku


  Mikael Niku             URL:
  University of Helsinki  Dept. Basic Veterinary Sciences

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