[Histonet] Re: bluing hematoxylin and alkaline water????

[Robert Richmond]

Hard water (water containing dissolved calcium carbonate and/or
sulfate is alkaline enough to use as a bluing agent by itself. New
York City, Hot Springs Arkansas, and San Antonio Texas - in my
personal experience - have tap water sufficiently alkaline that you
can blue hematoxylin in it in a reasonable length of time.

The original Scott's solution was in fact devised as a substitute for
alkaline tap water. Scott SG (Oxford). On successive double staining
for histological purposes—preliminary note. Journal of Pathology and
Bacteriology 1911-1912: 16,390-8. Scott notes that "As tap water
varies in constitution from place to place, and even the alkaline tap
water of Oxford from the oölite Cotswolds requires ten minutes with
occasional changes for safe removal of acid, the artificial substitute
mentioned above has been introduced." Robert Wyllie in 1970 told me
that this bluing solution was widely used in Australia, apparently
introduced by Oxonian histologists nostalgic for the tap water of
their homeland. Wyllie was an Australian histochemist who worked in
the pathology department at Johns Hopkins for a number of years. He
greatly simplified Scott's original formula, and referred to this
preparation as Scott's solution. Gary Gill popularized Scott's
solution along with his well-known hematoxylins.

Bob Wyllie (of blest memory) was an absolute genius in the
histochemistry lab, but he was a very modest man. He published very
little, and many of his ingenious techniques are totally lost (I have
just a few of them). I spent a year as a research fellow in his
laboratory in 1970.

Bob Richmond
Samurai Pathologist
Knoxville TN