Re: hazing H&E's

From:"MARY T HODGES" (by way of Histonet)

Ԫ     Pam  You are right on and additives to the tap water can really
play havic  on staining. Tere

   ----- Original Message -----    From:    To:       Sent: Tuesday, July 15, 2003 1:52  PM
Subject: Re: hazing H&E's
Pamela Marcum at Polysciences notes:

>>When I    lived in the Bay Area, Foster City CA, we found the tap water to at
pH 8.3    to 8.7.  If I tried to blue sections it was a joke so we gave up
just used the water as the bluing agent.  Now I am outside    Philadelphia and
the water is closer to pH 7.0 -7.8 and I need to blue    again.  so indeed
checking the pH of the water in your area is    important.  I found in my
travels as a technical person that the pH of    the water varies in all areas
of the country.<<

Hard water -    with dissolved calcium bicarbonate - has a high pH and will
hematoxylin quite effectively. I've seen this in New York City and in Hot
Springs, Arkansas, among other places. Scott's bluing solution began as an
artificial tap water. From my notes:

Scott SG (Oxford). On    successive double staining for histological
preliminary note.    Journal of Pathology and Bacteriology 1911-1912:
Scott notes    that "As tap water varies in constitution from place to
place, and
even    the alkaline tap water of Oxford from the olite Cotswolds
requires ten
minutes with occasional changes for safe removal of acid, the artificial
substitute mentioned above has been introduced." Dr. Robert G. Wyllie in
1970 told me
that this bluing solution was widely used in Australia,    apparently
by Oxonian histologists nostalgic for the tap water    of their homeland.
introduced this bluing technique at Johns    Hopkins, using the term Scott's
solution, and Gary Gill's publication    popularized both the term Scott's
and the solution    itself.

Scotts original formula (which Gary Gill simplified)    was

potassium bicarbonate 2.0 g
calcium chloride dihydrate 0.5 to    0.75 g
magnesium sulfate heptahydrate 20.0 g
water 1000.0 mL

Bob    Richmond
Samurai Pathologist
Knoxville  TN

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