Re: [Histonet] Large vessel endothelium problem

From:Gayle Callis


Next question:  Are you there when they slaughter of the animal, then 
collect the blood vessel as soon as possible after death? Or do you get the 
tissue after many hours of it sitting around in a slaughter house?  If not, 
you may be dealing with autolyzed tissue and  poorly preserved epithelium 
which may be sloughing off before you ever get a sample as per your 
assessment of "early steps of sample process."

You had written:
I need to do some immunostaining of large vessel endothelium
(paraformaldehyde-fixed paraffin material).
The problem is, the endothelium is easily stripped off and lost,
apparently in early steps of sample process.

This problem could be solved if you are there to collect and fix the tissue 
immediately after death of the bovine.  I suspect this problem could occur 
due to any delay in opening the vessel and delay of fixation after the 
death of the animal.  Be generous with the amount of fixative you use for 
the size of the tissue.

  At 01:36 AM 4/20/2006, you wrote:
>Hello again. Sorry for a bit unclearly formatted question.
>More specifically, I'm doing samples of bovine blood vessels, obtained 
>from a slaughterhouse. So perfusion is unfortunately out of question :)
>And this is about REALLY big vessels, such as the bovine aorta. So the 
>samples are bits (about 1 cm2) of vessel wall cut off and placed in PFA 
>for overnight at +4C, then processed for paraffin sections in a routine manner.
>And yes, this is primarily about blood vessels, not lymphatics.

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)

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