[Histonet] More details: decalcified sections of bone
I sent this out earlier (see below) and more details of my tissue processing
and sectioning were requested.
I want to identify apoptotic osteocytes in rabbit cortical bone as well as
some morphological features especially microcracks. I fix the bones in 4%
paraformaldehyde for 24 hours, then place then in 70% EtOH until I can
decalcify them in EDTA at room temp. Between the EtOH and EDTA steps I
rinse the bones in dH2O for about 3 days. After decalcification, the bones
are again rinsed in dH2O and then embedded in paraffin and sectioned to 6
microns. I use the TUNEL kit from Roche for In situ labeling of DNA breaks.
Do you think that I have to do decalcification for the TUNEL to work? Does
sectioning of paraffin embedded tissue create a lot of microscopic damage to
I hope this clears up my situation and goals.
>From: "Nicole Hedgecock"
>Subject: [Histonet] decalcified sections of bone
>Date: Tue, 05 Oct 2004 14:42:45 -0400
>I am attempting to do TUNEL staining of osteocytes in cortical bone. I was
>wondering why the bone has to be decalcified in order to do this process.
>And Why do the sections have to be so thin?
>Orthopedic Research Lab
>UCD Medical Center
>University of California, Davis
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