RE: decalcification in EDTA and nitric acid

From:Barry R Rittman

Myriam
Generally the only rationale for using glutaraldehyde in with the EDTA
is for electron microscopy. The disadvantage of using glutaraldehyde is
that it greatly prolongs the decalcification time. If you are
decalcifying using EDTA and the time is such that you fear maceration of
the tissues, you can always place the tissues back into fixative for a
short time. In general it is not wise to decalcify entire teeth for EM.
If this is for EM then nitric acid is contraindicated.
After nitric acid, thorough washing in water before processing.
Tests other than X-Ray, several chemical tests, such as using
calcein,are available however for intact teeth X-Rays are still the
method of choice as small amounts of mineral may not be detected by the
chemical methods. Chemical tests for decalcification rely on calcium
being present in the solution. As calcium is combined with EDTA and is
not readily available, I don't think that most mineral tests work with
EDTA. Please correct me someone of I am wrong.
Barry

-----Original Message-----
From: Myri37@aol.com [mailto:Myri37@aol.com] 
Sent: Wednesday, June 18, 2003 9:42 AM
To: histonet@pathology.swmed.edu
Subject: decalcification in EDTA and nitric acid

first of all thanks everybody for your precious help
i use EDTA 0.15 M with 1.9% glutarald#233#hyde in 0.06 M cacodylate
sodium buffer, after two months i put bone in nitrix acid 5 %, with 10%
glutarald#233#hyde for 3 weeks
I did switch from EDTA to nitric acid to increase decalcification rate
without demaging the tissu structure
Does anyone know technique to chek if decalcification is complete other
than X RAy? 
In case of Nitric acid decalcification, what do you use for rinsing to
eliminate acid before ethanol dry, clearing and paraffine embedding ?
thank you
Myriam 




<< Previous Message | Next Message >>