Re: [Histonet] GILL 3 Confusion

From:Gayle Callis

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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