[Histonet] Re: Question regarding 10% Neutral buffered

From:"John Auld"


It shouldn't make any difference to most immuno stains. The NBF in the EDTA
will continue to act as a fixative and may, just may overfix some Ags, some
will be OK.
I would suspect that 24 hrs primary fixation is insufficient prior to
decalcification anyway, so the use of NBF in EDTA maybe beneficial.
However, if some results have been obtained prior to changing the protocol
it maybe better not to change.



John Auld MSc CSci FIBMS
Dept of Histopathology and Clinical Cytology
Arrowe Park Hospital
Arrowe Park Road

Internal extn 2560
External Tel 0151 604 7025
Message: 25
Date: Fri, 10 Sep 2004 10:08:57 -0500
From: "Dunn-Jena, Patsy A" 
Subject: RE: [Histonet] Question regarding 10% Neutral buffered
To: "Histonet" 

Content-Type: text/plain;           charset="iso-8859-1"

Additional information.

The tissue is the palate/maxillary teeth of mice.  It is fixed in 10% NBF
for 24 hours prior to starting to decal in a 0.25 M EDTA which takes about
4 weeks.


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of
Dunn-Jena, Patsy A
Sent: Friday, September 10, 2004 9:49 AM
To: Histonet
Subject: [Histonet] Question regarding 10% Neutral buffered

I have a graduate student who has come to me with a question I cannot
answer as I have limited Immunohistochemical experience.  One of her
advisers wants her to go to using an EDTA decalcification recipe which
contains the 10% NBF in it which is a recipe we used on all of his previous
work.  She asked me about the NBF crosslinking her immuno sites as she has
been using an EDTA decalcification only recipe.  Some input here would be
nice so I can better explain it to her.

Patsy A. Dunn-Jena, RVT, LAT, HT (ASCP)
Mineralized Tissue and Histology Research Laboratory
Indiana University School of Dentistry
1121 W. Michigan Street, Room 238
Indianapolis, IN 46202

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