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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.
- [Histonet] BRDU, Tetracycline, calcein in bone Nancy W. Troiano
- Re: [Histonet] BRDU, Tetracycline, calcein in bone Robert Schoonhoven
