Re: [Histonet] Question about De-calcifying mouse paws
presumably you are facilitating fixative penetration by opening the skin?
You could try trimming the fixed sample to size by using a small bone saw,
we got one from this UK site, if you navigate you will come to some great
images of nasal turbinates in which the soft tissue is very well
preserved. We have used the same saw for LS sections of mouse femurs.
I also use a decalcifying agent which is formic acid based (plus
ingredients XXX) which does my whole fixed mouse heads or bone sawed head
slices of rat and guinea-pig in 24hrs. I also use it as a surface decal
of blocks if the teeth remain a bit solid. All of the immuno we have
done to date has worked as well as any tissues we have decalcified in EDTA
for much longer periods.
Price 139.90 Euros
Cat #1414 1 gallon
Quartett Immunodiagnostika - Biotechnologie GMBH
Schichauweg 16 - Berlin - Germany 12307
Fax:49 (030) 765-925-55
Contact Name Jürgen Gorczyza
GlaxoSmithKline Medicines Research Centre,
"Jamie E Erickson"
Sent by: email@example.com
[Histonet] Question about De-calcifying mouse paws
Here is my problem, We are a research histology lab which
supports groups doing Mouse/ Rat Collagen Induced Arthritis (CIA). The
researchers collected the paws and knees for routine processing (Paraffin)
and we take it from there. We are trying to give a quick turn around time
(2 weeks) on these studies so we are fixing the paws for 24 hours in 10%
neutral buffered formalin then switching it to CAL RITE (a
formaldehyde/Methanol/ Formic acid mixture from Richard Allan) for
decalcification. The paws sit in Cal-Rite for 2 days on a shaker and are
then trimmed by cutting the skin and one toe on each side off the paws or
trimming 1/3 off the knee Sagittal section (knee and some of the long
bones), this helps in decaling the paws and knee quicker. Then the samples
are put into fresh De-cal again for 2 more days before washing for 1 hour
in tap water and processing.
Problem: Knees are great , section great look great but some but not all
of the paws are chalky, white deposits in toes and ankles. This only
happens to about 1/3 of the
samples. I guess they are not de-calcified long enough?
Is there another way people are De-caling quickly with better results? Our
Pathologist is happy with the slides for the most part but the bad ones
are more difficult to section as you can imagine and sometimes the ankle
joints don't section well at all. Any Ideas would be greatly appreciated.
Sr. Research Associate
Abbott Bioresearch Center
100 Research Drive
Worcester, MA 01605-4341
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