You have been gifted with the most sensitive method, faster than chemical
endpoint determinations.  Lucky you!!!

You can now decalcify several bones in one container of acid decalcifier,
rinse and radiograph each one to determine if you have to go back into the
decalcifier. It is THE only method I use for endpoint determinations,
particularly when doing 50 rat knees at one time, piece of cake!!!  Bone
processing/sectioning and staining all improved using this endpoint
determination as overdecalcification was prevented.

There is an easier film to use, Kodak Diagnostic film, Ready-Pak, X-OMAT
TL, in 8 x 10 sheet contained in a sleeve so that you can work in a lighted
room until you need to develop the film.  Kodak Cat # 182 2089.

I tested endpoint with shelf at lowest level in Faxitron, 30 kvp for 1 min,
a manual operation.  If you need instruction manual on how to use the
machine, or buy parts, Faxitron Corp, 847 4659729 in Buffalo Grove IL 

Store your unused film in a freezer (no a self defrost) take out one, warm
to RT and use.  Your film will keep for years without loss of sensitivity.
You set up templates to identify each bone and can test more than one bone
at a time, make sure you view on an xray film viewer, a hand held magnifier
in a darkened room helps visualize tiny, residual calcium deposits.  

You set the bone on a sheet of zip lock baggie plastic, do not use Saran
wrap material,
put this on top of ready pak, shoot, etc, etc. BE sure you rinse the acid
from bone briefly, to avoid fumes, skin contact, etc. 

You can do unfixed or fixed predecalcified specimens to locate bone
lesions, etc.  Murine bones take only a few second to radiograph, and whole
mice and rats have been done in ours, sleeping peacefully.  

Have fun, this machine makes bone decalcification less of a chore, no more
quesswork on decalcification.  If the lab caught on fire, I would grab my
purse, Faxitron, Cryojane tape transfer, and run!!  

  

 

 
>Hello Histonetters:
>
>    What methods are being used to determine decalcification endpoints?  I 
>have inherited a Faxitran machine that uses Polaroid film and was wondering 
>if it could be used for endpoint determinations.  If so, any suggestions on 
>how to begin? (So far, I haven't even turned the darn thing on!!). Any input 
>is greatly appreciated!
>
>Diane Mahovlic
>Orthopedic Pathology Laboratory
>The Cleveland Clinic Foundation
>Cleveland, Ohio 
>
>
>
Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303