Re: Sectioning bone (REPLY)

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From:"P. Emry" <>
Date:Wed, 21 Apr 1999 12:50:48 -0700 (PDT)
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Thanks for the info.  I am wondering how the fat in the bone marrow is
preserved through the processing, what with clearing agents and alcohols.

What stains would you use to see the presents of fats in the marrow?

Thanks for your time.
On Tue, 20 Apr 1999 wrote:

> Dear Andrea:
> If you are looking at femoral head sections, remember that the decalcifying 
> process is a slow and tedious one.  However, with proper monitoring, the end 
> result can be fabulous.  I would recommend FIXING the specimen first in 
> formalin (depending on the size of the specimen) at least overnight.  We use 
> Surgipath Decal I, and change it every day.  It usually takes at least 4-5 
> days to decalcify a femoral head.  Change the solution at least once daily, 
> and rinse the specimen daily for at least 30 minutes in tap water (to remove 
> calcium salts).  When you finally can cut the specimen, and it is time to 
> process, make sure the tissue sections are no more than 2-3 mm thick.  
> After processing, embed the specimen diagonally in the mold.  This keeps you 
> from "facing" into too much of the specimen at one time.  I cut my blocks on 
> the vertical axis of the block holder on the microtome.  After gently facing 
> the block, rub a little warm water over the surface and slowly section the 
> block.  If it is still too hard, try immersing the block in cold decal 
> solution for 10-15 minutes after facing.
> We fix bone marrow biopsies in formalin for 2 hours, then in DECAL I solution 
> for 30-45 minutes.  RINSE WELL after decalcification.  Results have been 
> great.  (Again, I embed the specimen diagonally in the mold.)
> I know this is way too long, but bottom line I think the key to good bone 
> sectioning is fixation and changing decal solutions periodically.  
> Good luck with the bone!
> debra flynn
> jackson, ms  (sorry this was so long!)  :)

Trisha Emry 
U of Washington, Seattle

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