[Histonet] Re: Celestin blue B (was Help)

Mon Nov 7 11:55:06 CST 2011

Thanks Peggy for the helpful information about recent commercial haemalum substitutes. 

The MSDS for tango describes only an acidified solution of the dye (eriochrome cyanine R). This would need to be mixed with a ferric salt to make a blue nuclear stain. Iron-ECR is probably the best substitute for haemalum. See Dapson R, Horobin RW & Kiernan J (2010) Hematoxylin shortages: their causes and duration, and other dyes that can replace hemalum in routine hematoxylin and eosin staining. Biotechnic & Histochemistry 85(1): 55-63.

I agree with Peggy that newly blue and phoenix blue are probably iron-celestine blue, following the method of Lillie (Lillie RD, Pizzolato P, Welsh RA, Holmquist MD, Donaldson PT, Berger C (1973) A consideration of substitutes for alum hematoxylin in routine histologic and cytologic diagnostic procedures. Am. J. Clin. Path. 60: 817-819.), which uses two fairly stable stock solutions (iron alum and the dye) to make a working stain that keeps for a few weeks. Older formulations (Lendrum 1935, Gray 1956) contain the ferric salt, dye and sulphuric acid. They are more acidic (pH 0.8 to 1.1), keep for 6 months, and are also more selective for nuclei. Lendrum's stain is the one often followed by haemalum to give a black nuclear stain that resists acids (picric, phosphotungstic etc) even better than Weigert's iron-haematoxylin. If a company won't say what their product is, that's a good reason not to buy it. There's nothing new about celestine blue.

John Kiernan 
Dept of Anatomy & Cell Biology 
University of Western Ontario 
London, Canada 
 = = =
On 06/11/11, Lee & Peggy Wenk <lpwenk <@t> sbcglobal.net> wrote:

> 
> With the hematoxylin shortage of a couple of years ago (real, not imagined in about 2007-2008), several companies tried to come up with a synthetic dye substitute.
> 
> A little background: Celestine blue (CI 51050, also known as Mordant blue 14) is a substitute touted many years ago (late 1960s/early 1970s if I remember, when there was another shortage for different reasons). I've used it years ago, in a stain called MSB for fibrin (look in a Bancroft book). The MSB stain used a double nuclear stain of aluminum hematoxylin (e.g. Mayer or Harris) and celestine blue. We didn't like the celestine blue, because when we mixed the iron mordant salt with the celestine blue dye, it worked right away, but, the next time someone asked for a MSB stain (several months later), the celestine blue was overoxidized, and wouldn't work. So we would have to stop and make the dye up immediately. It was a nuisance. But the double nuclear stain to to try to keep the nuclei a blue color, after going through the next 3 dyes (MSB is sort of a quatrachrome). Eventually, we just started using the Weigert hematoxylin from the trichrome stain in place of the double nuclear stain in the MSB. Our pathologists like the MSB stain better with the Weigert hematoxylin.
> 
> Current use: Since 2008, three companies that I know of came up with hematoxylin substitutes, in response to that shortage.
> - Anatech Ltd, which used Mordant blue 3, CI 43820, and called it Tango Blue. So this is NOT celestine blue.
> http://www.anatechltdusa.com/MSDS_pdf/TangoStainMSDS.pdf
> - Newcomer Supply, which calls their substitute Newly Blue, but says it is a Celestine blue in one solution, ferric ammonium sulfate in the other solution. I don't know if you mix the two solutions before use, or if you dip the slides in first one solution and then the other. I didn't get around to their booth this year at NSH (sorry Marcia), and have never seen a procedure sheet on this stain.
> http://www.newcomersupply.com/products/standard-special-stains?page=N#181
> - ThermoFisher/Richard Allan, which calls their substitute Phoenix blue.The problem is, they are being secretive (proprietary) about what dye/reagents are in their solutions. I looked at one time, and couldn't find MSDS on their websites And their flyer doesn't mention the dyes name or CI #. However, the photo on the flyer, at least to me, looks like celestine blue. That's not proof that it IS celestine blue. Could be a close look-alike cousin.
> http://www.thermo.com/eThermo/CMA/PDFs/Product/productPDF_6906.pdf
> 
> So look to these companies for more information. Anatech has a newsletter, available on their website, about the hematoxylin shortage (and their Tango blue product, of course).  (Not that this will help you with your Celestine blue project, but it will fill you in on the hematoxylin shortage history.)
> http://www.anatechltdusa.com/Innovators/Inn08Summer.pdf
> 
> Hope that helps some.
> 
> Peggy A. Wenk, HTL(ASCP)SLS
> Schools of Histotechnology
> Beaumont Hospital
> Royal Oak, MI 48073
> 
> (Opinions expressed are my own, and not Beaumont Hospitals'.)
> 
> -----Original Message----- From: Bob Richmond
> Sent: Sunday, November 06, 2011 3:42 PM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Re: Celestin blue B (was Help)
> 
> Corrie Vernick writes:
> 
> >>I am currently a histology student at Keiser University. I am doing a project for my routine staining class about Celestine Blue. I've been able to find information on why it was created, the chemical make up, and some of it's uses including the trichrome stain. I am having trouble finding images of slides stained with Celestine Blue. Any additional information about the uses would be helpful as well! Thank you, Corrinne Vernick, Keiser University FL U.S.A.<<
> 
> I don't have access to my library this week, but you can get a good
> bit information by Googling celestin blue B. This dye was often used
> as a sort of backup or substitute for hematoxylin, particularly in the
> old outmoded Pearse stain for pituitary cells. R.D. Lillie as I
> remember didn't think much of the dye, and I don't think this dye is a
> very good topic for a study such as the one you describe.
> 
> Bob Richmond
> Samurai Pathologist
> Knoxville TN
> 
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