[Histonet] Mouse Paws

[Thomas Crowell]

Hello Histonetters,

I'm looking for some help or guidance in obtaining good sections from 
mouse front paws, to include the bones of the wrist and digits, as well as 
maintaining the surrounding matrix.  This is an ongoing problem in our 
laboratory and we have a difficult time obtaining good sections at this 
level, the problem appearing to be inadequate infiltration of paraffin 
into the interior of the paws, leaving the center somewhat mushy & white.

The feet are fixed in 10% NBF for up to a week, decalcified in formic 
acid/sodium citrate for 3 to 5 days, processed on a Shandon Pathcentre for 
45 minutes/reagent, 4 changes of paraffin, 45 minutes each, vacum & 
pressure on the waxes only.

Any suggestions regarding fixation, decalcification, or processing 
enhancements to improve the overall picture would be extremely  helpful.

Thanks!

Tom Crowell
BiogenIdec
Cambridge, MA