RE: Verhoeff's Van Gieson Stain-Aldehyde Fuchsin
|From:||"Monson, Frederick C." |
I have a protocol for using Gomori's Aldehyde Fuchsin (AF) with H&E to
insure an elegant stain of elastin in tissues in which your primary interest
is NOT in the epithelium - the AF does tend to stain GAG secretors which
confuses the issue. If I have to look/image GAG-secreting epithelium as
well as elastin, I would use Weigert's. I have not tried to block the GAG's
which might be an option.
My methods are similar in that either the AF or Weigert's is applied before
the H&E which is applied in normal fashion - I have not found the acid
differentiation of hematoxylin to adversely affect the AF or Weigert's.
An example of the AF H&E was to be found on the listserver's image page, but
I cannot check tonight as the site appears to be down.
Frederick C. Monson, PhD
Center for Advanced Scientific Imaging
West Chester University of Pennsylvania
Schmucker II Science Center, Room SS024
South Church Street and Rosedale Avenue
West Chester, PA, 19383
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From: NIDAL E MUVARAK [mailto:firstname.lastname@example.org]
Sent: Monday, March 03, 2003 5:11 PM
To: HistoNet Server
Subject: Verhoeff's Van Gieson Stain
Hi everyone. I just started doing VVG stains for mouse lung and artery
tissues. I did two trial stains. It seems that the length of the step that
requires differentiation in 2% ferric chloride is pretty crucial. In the
first experiment I incubated the slides in 2% ferric chloride for about 30
sec. The result was horrible staining.... the tissue was pitch black, with
lots of junk in the background. In the second experiment, I incubated the
slides for a little over 5 min. That resulted in the elimination of all
nuclei and elastic fiber staining (which should be black according to the
protocol). So I was wondering what's the optimal time for differentiation
with 2% FeCl3. I will list the protocol I'm using below. If anyone sees
anything wrong with it, please let me know. Thank you for your time.
1) Fix cryosections in cold aceton for 10 min. Dry. Rehydrate with distilled
2) Incubate in Verhoeff's hematoxylin (20ml alcoholic hematoxylin, 8ml 10%
FeCl3, 8ml Lugol's iodine) for 30 min.
3) Wash in tap water
4)Differentiate in 2% FeCl3. Check microscopically for black fibers on gray
5) Rinse in water
6) Rinse in 5% hypo solution for 1 min to remove iodine.
7)Wash in water.
8)Conterstain in Van Gieson's solution (1ml of 1% Acid Fuchin; 45ml of
saturated picric acid) for 5 min.
9)Rinse quickly with acidified water (5ml acetic acid in 1L distilled
10) Dehydrate 3X 95% EtOH, 2X 100% EtOH, 3X xylene. Cover with resinous
Nidal E Muvarak
Associate Research Specialist
Department of Biomedical Engineering
University of Wisconsin-Madison
1550 Engineering Dr.; Rm. 2158
Madison, WI 53706-1609
Lab: (608) 265-8921; Office: (608) 265-4205;
Home: (608) 256-7934; Cell: (608) 332-6068
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