[Histonet] BRDU, Tetracycline, calcein in bone

[Robert Schoonhoven]

You can also perform BrdU on the MMA sections the BrdU label is quite hardy.

Robert Schoonhoven




________________________________
From: Nancy W. Troiano <nancy.troiano <@t> yale.edu>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Tuesday, April 21, 2009 1:37:01 PM
Subject: [Histonet] BRDU, Tetracycline, calcein in bone

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. 

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