[Histonet] mice legs

Derek Papalegis derek.papalegis <@t> tufts.edu
Tue Sep 15 08:51:10 CDT 2009

Hi Sharon,
I frequently get whole mouse legs submitted to me and decalcification 
doesn't break down the tissue. I use Cal-Rite as a decalcifier but any 
formic acid decal would be sufficient. Since formic acid is pretty 
forgiving, I decal the legs for 48-72 hours on a rotator. I change the 
solution after 24-48 hours and then place them back in decal for 24 
hours. I have left bones in decal over the weekend and they have been 
fine. I have found that if you are focusing on the long bones, this is 
sufficient but if you are interested in seeing the ankles and feet, you 
should remove them from the leg and decal them longer. When processing, 
I use a longer program. Here is the program that I use:

70%    1 hr
80%    1 hr
95%    2 hr
95%    2.5 hr
100%  2.5hr
100%  3hr
100%  3hr
xylene 1hr20min
xylene 1hr20min
xylene 1hr20min
paraffin 1hr
paraffin 1hr
paraffin 1.5 hr
paraffin 1.5 hr

When processing, only process the bone samples with this program. If you 
batch other tissues in with the bones, the tissue will be over 
processed. Let me know if you have any more questions.


Derek Papalegis HT (ASCP)
Senior Histology Technologist
Division of Laboratory Animal Medicine
Tufts University 

Shaw, Sharon wrote:
> Good Morning Histo World,
> I would like to know if anyone is working with mice legs, I have a PI that I work with that wants to process the whole leg, the problem is I need to decal it first and is wondering if the decal will break down the tissue, I think it would he doesn't think so. And if anyone has do this would it be possible to share your protocol with me from decal to processing.
> Thanks,
> Sharon- WPI
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