[Histonet] BRDU, Tetracycline, calcein in bone

[Nancy W. Troiano]

I would recommend fixing the bones in 70% ethanol, embedding in 
methylmethacrylate undecalcified and cutting 8 - 10 micron thick sections 
for visualization of tetracycline and/or calcein labels.   Decalcification 
will remove fluorescent labels bound to the mineral in bone as mineral is 
removed by decal procedures.  You may need a larger microtome (Polycut) to 
cut these bones depending on the size.  The fluorescent labels in 
combination with a nice von Kossa stain would certainly show formation/ 
mineralization of bone in this area which you can quantify using an 
Osteomeasure or other system, however, you may want to do a few in paraffin 
for optimal BRDU label preservation.  I would recommend a slow embedding 
method for these bones - you can check our article in the JOH, June 2004, 
Vol. 27(2) pp119-130 which describes our slow embedding method for rat 
bones which can be adapted for rabbit bones.  You can contact me for 
suggestions and with questions if you would like to.