Marg,

In my experience, trouble with the Gordon and Sweet retic stain results from
several factors.  The making of the silver solution is critical, as too much
ammonium hydroxide can result in pale or weak results.  Also, if the tissue
section is very thin there will be less demonstrated reticulin.  If you save
your working silver solution to re-use and keep it in the refrigerator,
you'll either need to allow it to come to room temperature before using, or
extend the time you keep the slides in it if it's cold.  We have been able
to successfully re-use this solution for up to a week (depending on the
number of slides passed through there).  When in doubt, make it fresh.  :)
Also, is it possible the sections were over-toned in gold chloride?

We just recently modified our procedure with one that was working well at
another local hospital.  For what it's worth, here it is:

1.  Deparaffinize sections and hydrate to distilled water.
2.  Oxidize sections in 1.0% potassium permanganate solution for 5 minutes.
3.  Rinse in tap water.
4.  Bleach in 1% oxalic acid for 2 minutes, or until sections are colorless.
5.  Wash in tap water.
6.  Sensitize sections in 2.5% ferric ammonium sulfate for 10 minutes.
7.  Wash in several changes of distilled water.
8.  Impregnate sections with the working ammoniacal silver solution for 3
seconds.
9.  Dip each slide into 95% alcohol once and go immediately into the
reducing solution.
10. Reduce sections for 45 seconds in the formalin solution (2-4 drops
concentrated formaldehyde to 50 ml distilled water).
11. Wash in tap water.
12. Rinse in distilled water.
13. Counterstain in Van Gieson for 5 minutes.  Rinse in distilled water.
14. Dehydrate in two changes each of 95% and absolute alcohols, clear in
xylene, and mount with synthetic resin.

I do not have much experience with this modified procedure as we've only
been using it for about 2 weeks now, but our liver pathologist loves it.
The techs were (understandably) uneasy with the incredibly brief time in
silver solution, the weak formalin, and the omission of gold chloride and
sodium thiosulfate.  I asked them to follow the procedure to the letter, and
so far, so good.

Hope this helps,

-Teri Johnson, HT(ASCP)
Histology Supervisor
Physicians Reference Laboratory

----- Original Message -----
From: Hagerty, Marjorie A. <mhagerty@emc.org>
To: 'Histonet' <histonet@pathology.swmed.edu>
Sent: Thursday, April 06, 2000 4:06 PM
Subject: Retic Query


> Can anyone offer me any tips on the Gordon Sweets Reticulum?  I have done
a
> million and one reticulin stains, but not in a long time. When I looked at
a
> retic someone had done today, I would have liked to see more staining. Any
> little tips would be greatly appreciated.
>
> Thanks!
> Marg
> Supervisor, Anatomic Pathology
> Eisenhower Medical Center
> 39-000 Bob Hope Drive
> Rancho Mirage, CA 92270
>
>