Re: Verhoeff's Van Gieson Stain

From:aaj1@columbia.edu

Try acid orcein with methylene blue 1% counter stain.Quoting Geoff 
McAuliffe :

>     Carlos makes an excellent suggestion, stain elastic with
> aldehyde fuchsin. Easy to make, easy to stain, nice deep purple
> color, fool-proof differentiation. The paraldehyde used to make
> the stain must be fresh and is rather uncommon but can be found
> at a pharmacy. It is a controlled substance. Acetaldehyde can be
> substituted; I can dig up a reference if needed.
> 
> Geoff
> 
> NIDAL E MUVARAK wrote:
> 
> > 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.
> >
> > VVG Stain:
> > 1) Fix cryosections in cold aceton for 10 min. Dry. Rehydrate
> with distilled water.
> > 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 background.
> > 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 water).
> > 10) Dehydrate 3X 95% EtOH, 2X 100% EtOH, 3X xylene. Cover with
> resinous mounting medium.
> >
> > 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
> 
> --
> **********************************************
> Geoff McAuliffe, Ph.D.
> Neuroscience and Cell Biology
> Robert Wood Johnson Medical School
> 675 Hoes Lane, Piscataway, NJ 08854
> voice: (732)-235-4583; fax: -4029
> mcauliff@umdnj.edu
> **********************************************
> 
> 
> 





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