Andrea:

This technique may be adapted to suit your needs:

Humberstone's
Elastic stain

Humberstone's solution -

 Victoria blue 4R
                             1 g
 Ethyl violet
                             1 g
 Distilled water
                          200 ml
 Resorcin
                             4 g
 Dextrin
                            0.5 g
 Ferric chloride 30% (fresh)
                           25 mL
 Ethanol 95%
                          100 mL
 Phenol
                            10 g
 Concentrated hydrochloric acid
                            4 mL


Add the victoria blue and ethyl violet to the 200 mL water.
Boil to dissolve.
Use an oversize flask to avoid foaming.
Add the resorcin and dextrin
Add the ferric chloride.
Boil for 3 minutes, cool and filter.
Place the precipitate and filter paper back into the original beaker.
Add 100 mL ethanol.
Boil gently for 15 minutes.
Cool and filter.
Restore the volume to 350 mL with 95% ethanol.
Add the phenol and hydrochloric acid.
The solution is stable at room temperature. Stain intensifies with age.

Sections

Formalin or formal sublimate fixation is satisfactory.
Paraffin sections at 5u are suitable.

Method

   1.Bring sections to water via xylene and ethanol
   2.Do a Mallory bleach using 0.25% potassium permanganate for ten minutes.

   3.Rinse with 95% ethanol
   4.Place into Humberstone's solution overnight.
   5.Wash with 95% ethanol to remove excess solution.
   6.Wash in water.
   7.Counterstain with van Gieson. *
   8.Dehydrate with ethanol, clear with xylene and mount with a resinous
medium.

  *Omit van Gieson counterstain, if no collagen is to be stained.

Expected Results

     Elastic fibres - greenish blue black
     Collagen - red
     Cytoplasm - yellow
     Nuclei - blue

Reference

An Elastic Tissue Stain
Humberstone, G C W, and Humberstone, F D.
Journal of Medical laboratory technology, Vol. 26, No 2, pp. 99. (April
1969)

Eric Kellar
Histology/Immunohistochemistry
University of Pittsburgh Medical Center

	----------
	From:  andreah@imclone.com [SMTP:andreah@imclone.com]
	Sent:  Friday, December 04, 1998 5:20 PM
	To:  HistoNet@Pathology.swmed.edu
	Subject:  Vessel dye?? Part 2

	Well here is something I just found, Victoria blue elastic stain ...
does
	anyone know about this stain and whether or not it would stain
vessels
	adn not collagen??

	Andrea
	ANDREAH@IMCLONE.COM