Re: Intestine fixation
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From: | "J. A. Kiernan" <jkiernan@julian.uwo.ca> (by way of histonet) |
To: | histonet@histosearch.com |
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My two cents' worth (or ha'p'orth from under my British hat)
is based on some work done in my lab about 12 years ago.
We were collecting 2 cm pieces of large and small intestine
from dozens of rats & mice (mostly mice) that had been dosed with
senna and related purgatives. We needed to know the distances of the
samples from the pylorus (or from the caecum), and also the
total length of the small or large intestine (Both are both rather
variable because it stretches. With measured lengths, the position
of a sample can be identified as a proportion of the whole
length.)
To get a straight, measurable, whole GI tract, the simple answer
was a piece of plastic eavestrough (that's gutter in UK) about
5 feet (1.5 metres for Young Canadians) long. To use it, put one
end over a sink and elevate the other end about 6" or 15cm. Lay
out the intestine along the length of the trough, with the anal
end over the sink. (Remove the caecum and treat the small
intestine and colo-rectum as separate, parallel objects.)
Straighten out any loops & twists and make the length measurements.
Then inject saline from a 20 ml syringe, downhill into the sink.
The point I'm trying to make is that a length of plastic gutter
or eavestrough (mine is still quite brightly white and stands
in a corner taking up hardly any space) is a great petty-cash
item (about $5) for any lab that has to handle and clean out the
intestines of small animals. Imagine what a lab supply company
might charge for a "fully adjustable oblique-flow intestinal
irrigation apparatus, customized for small rodents."
John A. Kiernan,
Department of Anatomy & Cell Biology,
The University of Western Ontario,
LONDON, Canada N6A 5C1
P.S.
The saline should be followed by fixative if you need well
preserved villi. Nearly all our material, however, was made
into whole-mounts containing the myenteric plexus,
which was supposedly susceptible to the metabolites of senna.
If you have read this far and want to know that purgatives do
very little to mice and their enteric neurons, please email me
personally and I'll send you a reprint. In addition to the senna
study, we did neuron counts in wholemounts stained for RNA with
cuprolinic blue. This method has since been used, by others, for
larger animals, including cattle.
J.K.
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