There is a question on the validity of measuring osteoid on thick sections.
The reason is that it is rare in bone to have sections where edges are absolutely vertical throughout the section. You therefore have several planes superimposed and several edges in these planes resulting in a penumbra effect. The greater the angle to the vertical edge of the bone such as edges of trabeculae or Haversian canals, the greater the errors in measuring.
The most accurate measurements are those utilizing sections that are 5 microns or below.
Is there some particular reason that you must use sections this thick?
Barry

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Herrick, James L.
Sent: Tuesday, September 16, 2008 5:12 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Question on staining

Hi all,

I am trying to stain for osteoid and bone in pig tibia (need to quantify osteoid and bone volumes). Does anyone have experience with these types of stains in 40-60 µm thick, MMA embedded, sections? I have tried a Masson's trichrome and Toluidine blue, but have been unable to get well defined stain differentiation with this thick of sections. I would really appreciate any help I can get. Thanks again.

Jim

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