Re: [Histonet] fixing rat tibia

From:Gayle Callis

70% alcohol fixation to preserve the tetracycline - bones can sit in this 
for several days, and I would opt for longer time due to bone 
density.  People also use alcoholic formalin which does not remove the 
fluorescent label.  After fixation is complete, you should transfer bones 
from this fixative into 70% fixative.  The alcoholic formalin improves 
morphology for other staining from the same block.  If you use this, 
Anatech has a superb commercial formulation, and to test how long it takes 
to fix bone, work with several tibias  (unlabeled) and open one sample 
every day for 1,2,3,etc days.  If the bone is still bloody inside, then you 
need to fix longer, it should appear grayish, as formalin fixed tissues 
would appear when well fixed.

You should not have to bisect the rat tibia. That is difficult to do and to 
avoid shattering bone, you could purchase a little hobby saw, like a Dremel 
rotating tool/hand saw, but you need to find one that has a tiny little saw 
blade for cutting.    Unfortunately, the small  saw blade for Dremel was 
discontinued, probably for safety reasons, but look for a similar blade at 
a hobby shop or a similar handtool setup.  You may find a blade that fits 
the Dremel tool.

Surprisingly, hobby shops for woodworking, etc often have minitools 
suitable for bone work, just wear eye safety wear, things tend to fly 
around and spray blood and bone dust, very messy.

Good luck

  At 06:14 AM 4/26/2006, you wrote:
>    Could anyone advise me regarding the time required to fix a rat
>tibia. Can it be fixed whole or is it best to slice in half longitudinally.
>Also is 70% ethanol appropriate as a fixative for bones that have
>been labelled with Oxytetracycline. Another dilema I have is that I
>dont have access to a specialist tools, should I need to bisect the
>tibia in half. What tools could be used to cut the tibia in half
>longitudinally without cracking the bone. I am new to all this so any
>any help / suggestions will be appreciated. Thanks
>                              Ash
>M13 9PT
>TEL:+44 (0)161 275 5177
>FAX:+44 (0)161 275 5958
>Histonet mailing list

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)

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