[Histonet] Re: RED COUNTERSTAINS

Fri Feb 29 18:01:33 CST 2008

Actually, I tend to run about 5 experiments at the same time so if I have to leave something for 30 minutes, I usually welcome the reprieve <grin>.  Also, I was just getting such inconsistent results with nuclear fast red and it was irritating me <more grins>.  Having worked previously in Biotech and working currently in academia, I've discovered that what you won't tolerate in one, you will in the other and vice versa.

Jacqui

Jacqui Detmar, Post-doctoral Fellow
Samuel Lunenfeld Research Institute, room 876
Mount Sinai Hospital
600 University Avenue,
Toronto, ON, Canada
M5G 1X5

Tel:       416-586-4800 x2451/x2290
Fax:      416-586-8588
email:   detmar <@t> mshri.on.ca

________________________________

From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Houston, Ronald
Sent: Fri 2/29/2008 1:12 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Re: RED COUNTERSTAINS

Can't think why you would want to wait 20-30 minutes for a nuclear
counterstain

Ronnie Houston, MS, HT(ASCP)QIHC
Anatomic Pathology Manager
Nationwide Children's Hospital
700 Children's Drive
Columbus, OH 43205
(614) 722 5465
Ronald.Houston <@t> NationwideChildrens.org
Columbus Children's Hospital is now Nationwide Children's Hospital
www.NationwideChildrens.org

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jacqui
Detmar
Sent: Friday, February 29, 2008 12:39 PM
To: Geoff McAuliffe; Robert Richmond
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Re: RED COUNTERSTAINS

Hi all.  I also bought some brazilin powder (from Anatech) and mixed it
according to directions, and the initial results were disappointing.  I
then prepared an alum brazilin solution (recommended by someone on
Histonet, I think, but I'm at a conference right now and don't have
access to my e-files! <grin>), which is pretty much the same way you
would make alum hematoxylin, except you substitute brazilin for
hematoxylin.  I remember having to use slight heating to get some of the
powder into solution, and even then, not all of the brazilin went into
solution (it falls to the bottom...I guess you could filter it, but I've
never bothered). 

I find that I get the best results if I stain my slides for 20-30
minutes in the alum brazilin, followed by tap-water washes and a final
"blue-ing" in Scott's blueing solution for 30 seconds.  The result is a
nice, raspberry-coloured stain.  The recipe for the brazilin solution is
in the sheet given by the manufacturer.  I love this stain and I no
longer use nuclear fast red.  The only time I've had trouble with it is
with the von Kossa stain.  You *cannot* use brazilin after von
Kossa...the stain goes away for some reason...you need to use methyl
green or nuclear fast red.

I think that's about it!  If you have any questions, please feel free to
ask!

Jacqui

Jacqui Detmar, Post-doctoral Fellow
Samuel Lunenfeld Research Institute, room 876
Mount Sinai Hospital
600 University Avenue,
Toronto, ON, Canada
M5G 1X5

Tel:       416-586-4800 x2451/x2290
Fax:      416-586-8588
email:   detmar <@t> mshri.on.ca

From: Geoff McAuliffe
Sent: Fri 2/29/2008 10:25 AM
To: Robert Richmond
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Re: RED COUNTERSTAINS

Hi Bob:

    I bought some Brazilliant recently and mixed it according to the
directions provided. The results were dissipointing, red-orange nuclei,
not very intense. The tissue was rabbit kidney fixed in
formalin-alcohol-acetic. An alum hematoxylin on the same tissue looks
fine.

Geoff

Robert Richmond wrote:
> Diana McCaig asks about the nuclear counterstains nuclear fast red and
> neutral red.
>
> Does anyone on this list have any experience with Anatech's
> "Brazilliant", their trade name for alum brazilin, closely related to
> alum hematoxylin, but red instead of purple? This looks to me like a
> very logical red nuclear stain, and I'd certainly like to see it in
> action if it were possible for me to obtain it (remember that hospital
> pathology services are not usually permitted to order from small
> companies like Anatech and the Davidson marking ink people).
>
> As everybody on this list ought to know, hematoxylin is a dye
> extracted from the logwood tree (Haematoxylum campechianum), with an
> aluminum mordant. (There is no satisfactory synthetic substitute.)
> Brazilin is structurally very similar, but with an alum mordant it is
> red rather than purple. Brazilin is extracted from the closely related
> brazil woods, Caesalpinia echinata or C. sappan.
>
> One would expect this red dye to have the same staining specificity as
> hematoxylin, and it should not wash out in aqueous mounting media.
>
> (I have no connection with Anatech.)
>
> Bob Richmond
> Samurai Pathologist
> Knoxville TN
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>  

--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff <@t> umdnj.edu
**********************************************

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