Re: hazing H&E's
Dear Bob and All,
Here in Australia I have "grown up" on, and still use, a solution called
Scott's Tap Water Substitute.
The tap water here in Townsville is usually pH 5.5, so we need something to
make Haematoxylin go blue.
Scott's TWS is pH 9 and does the job in 30seconds, and does not tend to
lift sections like more alkaline solutions.
The formula given in the 4th edition of Carleton's Histological Technique is;
3.5 gm sodium bicarbonate
20gm magnesium sulphate
thymol to inhibit mould.
The authors quote Scott in the same article that Bob mentioned.
At 01:52 PM 07/15/03 -0400, RSRICHMOND@aol.com wrote:
>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 and
>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 blue
>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 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,
>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." Dr. Robert G. 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
>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.
>Scott’s 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
Mr.Laurie Reilly Ph 07 4781 4468
Physiology & Pharmacology Fax 07 4779 1526
Aust.Inst.of Tropical Vet.& Animal Sc.
James Cook University
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