Re: Osteocyte fixation

From:Connie McManus <>

Fixatives for osteocytes:

If you're doing this in paraffin, I'm assuming you are decalcifying your bones.
The length of time and the type of decalcifying agent will affect the cellular
details in your sections.  I would suggest using 10% EDTA to dacalcify.  If left
for too long, acid decalcification (such as nitric acid or formic acid formulas)
can destroy the nuclear detail in the cells making them appear washed out.  On the
other hand, EDTA takes a long time to do, but it's worth it.  I'm currently using
it on fish heads in a whirling disease project and the nuclear details are crisp
and sharp.  The EDTA also makes sectioning easier.  If you go to the histo
archives, there was a good discussion on this a couple of months ago with some
good recipes.

Connie McManus

john baker wrote:

> This is for one of our graduate students here in the Orthopaedic Research Lab.
> What is a good fixative to preserve osteocyte morphology in bone biopsies?
> The biopsy is from regenerated bone filling in a  defect in a rat femoral
> cortex.  The biopsy is maintained in tissue culture for several days then
> embedded in paraffin, sectioned, and stained with Alcian blue Hematoxylin.
> The purpose of the study is to acertain the viability of the osteocytes.
> If you know of a good way to demostrate cell viability within tissue please
> include that too.  If you require more information to help answer this
> please let me know.
> Thank you,  John  for Ed Hoffler
> John A. Baker
> The University of Michigan
> Orthopaedic Research Laboratories
> Histology Unit
> 400 North Ingalls, G161
> Ann Arbor, MI 48109-0486
> my office 734-936-1635
> lab office 734-763-9674


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