[Histonet] Mouse Joints

[gayle callis]

Dear Becky, 

You didn't say what decalcifier you were using?     It is a good idea to
test the decalcification endpoint on these bones before processing .   I
have a very easy weight loss/weight gain method to make sure the calcium is
removed from these joints or any bone for that matter.    We used this for
murine knees, all long bones and paws and never had problems with residual
calcium.   

Two things: 

1.  Now that the bones are embedded and contain calcium, you can surface
decalcify by immersing the trimmed block face into any acid decalcifying
solution, particularly one from a vendor,  for a few minutes.  This will
remove the calcium from only a few um of the sample, so be careful to NOT
trim this minimally decalcified portion away when you resection the block.
Be sure to rinse the acid off the block or metal parts on the microtome will
corrode.  You can re-cool the block after surface decalcification.   This
will take time for each block as you would have to repeat this each time you
want to cut further into the sample.  

2.  If you have a tungsten carbide knife, you could try sectioning with the
TC knife,  and not surface decalcify.   

Good luck 

Gayle Callis
HTL/HT/MT(ASCP) 

 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Berger,
Rebecca
Sent: Friday, February 14, 2014 9:07 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Mouse Joints

Hey Histonet!
 I have to troubleshoot some mouse knee joints that were not completely
decalcified before processing. I received them already embedded and they're
almost impossible to cut. Any tips to get them decalcified?
Thanks!
Becky
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