I think Sigma, or whoever wrote the specifications are uninformed. The
differences between Gill (progressive) and Harris (regressive) is discussed
in most modern histotechnology textbooks including Sheehan and Hrapchak's
book. Gill 1,2, or 3 are progressive and there have been some delightful
spinoffs on this formulation for excellent H&E staining.
Publication is Gill G et al. A new formula for a half oxidized hematoxylin
solution that neither over-stains nor requires differentiation. Acta
Cytologica, 18:300-311, 1974. The title itself indicates this is NOT a
regressive hematoxylin. When commercial Gill first came out, we bought ours
from Lerner Laboratories before everyone started to make Gill 1, 2, and 3,
and I still have the spec sheet on file.
Staining is done in 1 - 5 minutes, rinse, 4% acetic acid rinse 10 dips, tap
water rinse, Bluing reagent (Scotts Tap water, or Richard Allan Bluing
solution, pH 8) 10 dips (longer is permitted) then water rinse.
Some people will decolorize Gill hematoxylins. However, Gills publication,
and another by Meloan and Puchtler titled Harris Hematoxylin, What Harris
really wrote and the mechanism of hemalum stains. J Histotechnology
10(4):257, 1987 - you learn Gills do not need decolorizing per se with an
acid/alcohol. I think one reason people tend to decolorize Gill is they
stain too long in the solutions or use the more concentrated Gill 3 when
Gill 2 will do the job. We always tried to control for optimal staining by
time in the Gill rather than removing it later. Our staining time was
usually 1 1/2 to 2 minutes in Gill 2.
I am forever thankful to Gary Gill for making my hematoxylin and eosin
staining "life" easier!! He also has made comments on Histonet, so check
out the archives.
Gayle Callis HTL, HT, MT(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717
PuchAt 01:15 PM 5/2/2007, you wrote:
>I'm making this follow-up post because I'm getting some conflicting
>information about Gill 3 Hematoxylin. Quoting from the Sigma 2006-2007
>Biochemicals, Reagents & Kits catalog, page 1196: "General purpose
>nuclear stain, regressive type. Used with hematoxylin and eosin
>staining." Thus my question about substituting Harris' for Gill 3.
>I've had a couple responses telling me that Gill 3 is PROGRESSIVE. So,
>Sally Breeden, HT(ASCP)
>NM Dept. of Agriculture
>Veterinary Diagnostic Services
>PO Box 700
>Albuquerque, NM 87106
>Histonet mailing list
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