>Date: Wed, 25 Nov 1998 15:17:18 -0500
>From: Jeff Silverman <peptolab@hamptons.com>
>Subject: Re: cold hematoxylin
>To: Ian Montgomery <ian.montgomery@bio.gla.ac.uk>,
>        HistoNet@pathology.swmed.edu
>MIME-version: 1.0
>
>Go with Gill 3- it never needs filtering, stains frozens in 20-30 seconds,
>counterstains immuno's in 30-60 seconds, clean and crisp chromatin, stable
>and non-toxic, can't overstain etc. etc. Six minutes works nice in our H&E
>and 1minute in our Pap stain.  Differentiate in 1% HCl in etoh for paraffin
>and blue in dilute ammonia, for frozens blue only- no acid, and for cyto's
>1 minute and differentiate with  .5% aqueous HCl followed by 1 min water
>rinsse to blue. Look for the start of pink runoff in acid baths and stop
>immediately with water rinse. Works great every time and lasts 2-sometimes
>3 weeks in the staining bath.
>Jeff Silverman
>Southampton Hospital NY U$A
>peptolab@hamptons.com
>

Jeff,
	Never actually used Gill3, stick with 1 and 2. Normally use Mayer's
haematoxylin for 5 minutes then blue in Glasgow tap water, fresh from Loch
Katrine, for another 5 minutes. Frozens, couple of minutes, immunos, 20
seconds. No differentiating in 1%HCl/Alcohol or blueing in dilute ammonia.
The result, just as my wife describes me, "wonderful, just wonderful".
Ian.

Dr. Ian Montgomery,
West Medical Building,
University of Glasgow,
Glasgow,
G12 8QQ,
Scotland.
Tel: 0141 339 8855 Extn. 6602.
Fax: 0141 330 4100.
e-mail: ian.montgomery@bio.gla.ac.uk