Decalcifying mouse feet
|From:||Gayle Callis <firstname.lastname@example.org>|
First of all, make sure the feet are fixed. They contain so many tiny
bones, it takes longer than you think, I would fix a week to insure it is
complete fixation with NBF.
For enzyme staining, it would be best to do undecalcified bone frozens, not
always possible, EDTA inactivates alk phos, but you activity is restored
tactivity with magnesium chloride (Mg is chelated by EDTA). Acid
decalcifiers will probably destroy the enzyme, formic, HCl, any of these.
Decalcification with EDTA will take a long time, and you should try and
check it with xray to insure calcium is removed.
Hrapchak and Sheehan Theory and Practice of Histotechnology has excellent
discussion of this procedure, p 95 in second edition.
At 09:41 AM 1/19/01 -0800, you wrote:
>Thanks to all who replied to my TRAP question. I have another one: How do
>you prepare the mouse feet for the staining? What do you use as decal (we
>currently use 10%NBF)? How long do you decal the feet?
>Palo Alto, CA
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
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