[Histonet] Question about De-calcifying mouse paws

From:"Jamie E Erickson"

HI All,
           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.


Jamie Erickson
Sr. Research Associate 
Department: DSMP
Abbott Bioresearch Center
100 Research Drive
Worcester, MA 01605-4341
FAX: 508-793-4895
e-mail: jamie.erickson@abbott.com
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