Re: Mayer's hematoxylin and chloral hydrate

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To:Phyllis Davie <>
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>I have been having trouble with the hematoxylin counterstain for my
>immunos.  It is generally weak, with poor nuclear detail.  I have been
>purchasing Mayer's hematoxylin, and I have tried 4 different vendors now,
>all with the same results.  My first question is; what can be causing
>this weak staining, and what can I do to fix it?  (I have been
>counterstaining from 4-15 minutes, and many times, the staining--poor as
>it is--drops off markedly after just a few racks of slides)
>The second part of my question is this;  We decided to make our own
>Mayer's hematoxylin, but the recipe we found in the AFIP Laboratory
>Methods in Histotechnology, p. 52, calls for 50g of chloral hydrate.
>Unfortunately, chloral hydrate is a class IV DEA (Drug Enforcement
>Agency) regulated substance, and no-one here has a DEA license.  Can
>anyone tell me what is the purpose of the chloral hydrate in the
>hematoxylin is?
>Thank you
>Phyllis Davie
>Clinical Laboratory Supervisor
>PhenoPath Laboratories
>Seattle, WA
>(206)374-9009  phone
>(206)374-9009  fax

Dear Phyllis,

The recipe we use for Mayers Haemalum is as follows:-

Haematoxylin                       1g
Sodium iodate                      0.2g
Potassium alum                    50g
Distilled Water                      1 litre

Dissolve the haematoxylin in distilled water using gentle heat.Add the
Potassium alum and again use heat to dissolve it.When the alum has
dissolved and the solution has cooled add the Sodium iodate and shake
well.Filter the solution and add 20ml glacial acetic acid.

We have been using this recipe for many years without any problems.

Hope this helps,

Best wishes,

Tony Brandwood,
Dept. of Veterinary Pathology,
University of Liverpool,
P.O. Box 147,
L69 3BX.

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