RE: [Histonet] Re: Hematoxylin

From:Gary Gill

Hello Bob:

Gill hematoxylins more closely resemble Mayer's than Harris's hematoxylins
in terms of hematoxylin concentration (2 gm/L [gill] vs. 1 gm/L [Mayer {5
gm/L for Harris}]) and iodate oxidizer (Harris uses mercuric oxide).
However, Gill's No. 1:

*	Uses half the sodium iodate to oxidize the hematoxylin (0.1 gm/gm
hematoxylin vs. Mayer's 0.2 gm) to compensate for the variable amount of
hematein ordinarily present in hematoxylin (oxidized hematoxylin becomes
hematein, which combines with the Al mordant to form a "lake" that colors
chromatin [hematoxylin alone does not impart color]), and therefore, avoid
overoxidation.  Overoxidation produces oxyhematein, which is brownish, and
shortens the useful life of a hematoxylin formulation.

*	Uses glacial acetic acid, rather than citric acid.  Glacial acetic
acid separates some of the aluminum ions from the hematein molecules to
bring the working concentration of Al-hematein to approximately the same
starting concentration each time a batch is prepared.

*	Uses 1 part ethylene glycol (antifreeze) to 3 parts water to
stabilize the solution and to increase the solubility of the solution for
Al-hematein.  In Harris formulations, a greenish-golden surface precipitate
forms on standing.  This precipitate is sometimes identified as an
overoxidation product.  In fact, it is the "good stuff" -- pure Al-hematein.
It forms because the ingredient hematoxylin continues to oxidize over time
as the surface of the solution is exposed to atmospheric oxygen.
Eventually, the concentration of Al-hematein thus formed exceeds its
solubility limit in water, and so it precipitates.  It won't precipitate,
however, in a 25% ethylene glycol solution.

The greenish-golden surface precipitate can be retrieved by filtering the
Harris hematoxylin solution, and redissolved in 25% ethylene glycol.  The
resulting solution resembles a diluted hematoxylin solution.  It stains a
test buccal smear just as though it were full strength hematoxylin.  Ergo,
the precipitate is not junk, but the "good stuff".

The late Irwin Lerner registered my surname with the US Patent and Trademark
Office in 1973 without my permission.  That trademark has expired, so anyone
can use the Gill name with my formulation.  My permission was not, and is
not, required.  In the mid-70s, Lerner was sued to block some companies
(e.g., Polysciences) from using Gill in conjunction with my formulation.  I
assisted defense attorneys in fighting the suits.

BTW, I receive no royalties for this product.  However, I'm always open to
potential offers.

Gary Gill 

-----Original Message-----
From: RSRICHMOND@aol.com [mailto:RSRICHMOND@aol.com] 
Sent: Thursday, January 29, 2004 2:20 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Re: Hematoxylin


Mark Lewis, Product Specialist, Anatomical Pathology, Clinical Diagnostics, 
Thermo Electron Corporation asks us:

>>I'm taking a survey to find out which type of Hematoxylin is most 
>>commonly
used in Histology and Cytology labs.<<

And Gary Gill responds: >>I recommend Gill's for everything.  ;-)

The Gill hematoxylins are iodate-oxidized (Mayer) hematoxylins. I believe - 
please comment on this, Gary - that the term "Gill hematoxylin" is
proprietary 
and cannot be used without Gary Gill's permission.

The term "Harris hematoxylin" is correctly applied only to hematoxylins 
oxidized with mercuric oxide. Since mercury is a hazmat that (usually) is
not 
permitted in present-day histology labs, the term "Harris hematoxylin" has
come to 
mean any alum hematoxylin that the Marketing Department decides to call
Harris 
hematoxylin. This usage is deplorable. Iodate-oxidized hematoxylins should 
not be called Harris hematoxylin.

Bob Richmond
still the Samurai Pathologist
Gastonia NC
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