[Histonet] Trouble shooting decalcified bone sections (paraffin embedded)

[Kristen Lauing]

I section EDTA-decalcified mouse tibias frequently, and I find it helps if the paraffin is very very cold during sectioning - if the tissue starts shredding again, I press a large piece of ice to the block for a minute to cool it down without having to remove the block from the microtome.  It seems to work well for me, although it may not be the most technically sound way to get the job done.  I can usually get a good ribbon of sections this way when I'm having difficulty with a particular sample.
I have to use an old microtome and blades because I'm a student at a research institution, without the help of professional histotechs.  Our tibias are decalcified in 10% EDTA for 7 days at 4 degrees with agitation with frequent solution changes.

Kristen

>>> "CHEN, YIJING" <ychen9 <@t> kent.edu> 09/12/10 12:50 AM >>>
Hi,
We are having difficulty sectioning paraffin-embedded decalcified adult
mouse autopods (paws).  The tissues shatter as soon as they hit the blade
when sectioned.  

The autopods were soaked in CalEX for 4 days at room temp and felt extremely
soft before embedding, suggesting effective decalcification.  We use the
Sturkey EXTREMUS low profile disposable knives on our microtome.  These
knives are said to be coated with the hardest nitride coating available.
Any suggestions will be greatly appreciated.
Sincerely,
Yijing


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