[Histonet] Re: RED COUNTERSTAINS

[Jacqui Detmar]

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
>
> _______________________________________________
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> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>   


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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
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