[Histonet] Bone labelling for Microdamage

Jack Ratliff ratliffjack <@t> hotmail.com
Tue Dec 1 11:36:13 CST 2009


If you are looking for microdamage or are using basic fuchsin, I have  
been taught to bulk stain with 1% basic fuchsin in your processing  
solutions (i.e. 1% BF in 70% EtOH, 1% BF in 80% EtOH, 1% BF in 95%  
EtOH, and 1% BF in 100% EtOH) and then finish as you would with the  
remaining steps for your MMA protocol (i.e. xylenes, MMA +DBP, etc.)  
to achieve a fully polymerized bulk stained specimen. You then cut,  
grind, and polish as required for use with the EXAKT system. I was  
told the reason you process with the BF is because the cutting and  
grinding causes additional microdamage and if you stain first the  
additional damage will not be stained.

If you are talking about labeling bone for calculations of BFR or MAR  
and the animal did not receive timed injections of calcein, alizarin,  
tetracycline, or some other fluorescent bone label before necropsy,  
then you could try doing a 0.6% FA etch for 30 seconds followed with  
Sanderson's Rapid Bone Stain for 3 minutes, brief tap water rinse, and  
blot dry.

What is the exact specimen you are working with and what do you  
specifically wish to demonstrate????


On Dec 1, 2009, at 10:04 AM, Karie Reaser <kreaser <@t> vet.upenn.edu> wrote:

> Is it ok to process cortical bone, embed in MMA, section and grind  
> on the Exakt system. Last do the bone labelling stain? Or do I have  
> to do the bone labelling stain prior to embedding, sectioning and  
> mounting on slides? Any suggestions would be greatly appreciated.
> -- 
> Karie L Reaser A.S.
> New Bolton Center
> University of Pennsylvania
> School of Veterinary Medicine
> Comparative Orthopedic Research Laboratory
> 382 W Street Road
> Kennett Square, PA. 19348
> Phone:610-925-6278
> Fax:610-925-8120
> Email:kreaser <@t> vet.upenn.edu
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