RE: H & E Staining Method

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From:"Gary W. Gill" <>
To:Stan Harvey <>, mosborne@UNMC.EDU,
Date:Sat, 04 Sep 1999 10:53:06 -0500
Content-Type:text/plain; charset="iso-8859-1"

Mayer's is the least strong of all the surviving hematoxylin formulations.
Introduced in 1903, Mayer's formula included 200 mg per gm hematoxylin.
While that amount would be stoichiometric, (i.e., quantitatively just right)
if the hematoxylin were pure, hematoxylin is never pure.  A natural product,
it contains hematein, oxyhematein, and who knows what else.  Bottom line,
the amount of sodium iodate used by Mayer is greater than the amount
required by the amount of hematoxylin actually present so that there is some
overoxidation and less hematein present than expected.  The net result is an
even weaker Mayer's hematoxylin solution.  Ergo, trouble.

If you're using readymade Mayer's, this may be what you're running into.  If
you're making Mayer's, reduce the sodium iodate to 100 mg per 1 gm
hematoxylin.  In either case, you don't have the control you'd like and
you're using a very weak hematoxylin.  Increasing the staining time may not
solve your problem, as the concentration of stain within the tissue can not
exceed that in the solution.  BTW, there is no compelling reason to include
chloral hydrate in the formulation.  Indeed it is a controlled substance
(the stuff of knock-out drops, also known as a "Mickey Finn") and you may
not be able to buy it.  A pharmacist can.

Try using Gill's Hematoxylin No. 1.  It's an evolved Mayer's formulation
that includes among its modifications some described by Baker in 1962.

Gary W. Gill

-----Original Message-----
From: Stan Harvey []
Sent: September 01, 1999 9:01 AM
To: mosborne@UNMC.EDU;
Subject: Re: H & E Staining Method

I am using mayers and we are experiencing difficulties. The stain is too
I am looking for different methods of making up mayers.


Stanley Harvey
South Africa

>From: mosborne@UNMC.EDU
>Subject: Re: H & E Staining Method
>Date: Tue, 31 Aug 1999 13:49:58 -0500
>Did you try Mayer's hematoxylin?  There is no differenitation necessary.
>    Just my two cents worth.  Marianne Osborne

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