[Histonet] Question about De-calcifying mouse paws

[gillian.2.brown <@t> gsk.com]

Jamie,
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.

www.materials-science-nw.co.uk

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
Phone: 49(030)765-925-0 
Fax:49 (030) 765-925-55 
Website: http://www.quartett.com 
email: quartett <@t> t-online.de 
Contact Name  Jürgen Gorczyza


Regards

Gill Brown

GlaxoSmithKline Medicines Research Centre,
STEVENAGE,
Hertfordshire. UK
 



"Jamie E Erickson" <jamie.erickson <@t> abbott.com> 
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
17-Nov-2005 19:46
 
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Subject
[Histonet] Question about De-calcifying mouse paws






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.

Thanks

_______________________________
Jamie Erickson
Sr. Research Associate 
Department: DSMP
Abbott Bioresearch Center
100 Research Drive
Worcester, MA 01605-4341
508-688-3134
FAX: 508-793-4895
e-mail: jamie.erickson <@t> abbott.com
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