If you want to use the reagents you have already made up, try the following.  I usually do this differentiation step in 3-4 quick dips in the differentiating solution.  30 seconds would probably remove all your staining.  You can also put your slides back in the staining solution if you remove too much.  Check your slides under the microscope during the differentiation step until you are comfortable with the procedure.

Claye

>>>  03/11/03 11:21PM >>>
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 
> **********************************************
> 
> 
>