Orcein-Hematoxylin in Iodized Ferric Chloride as a Stain for Elastic Fibers, With Metanil Yellow Counterstaining

Abstract

Autopsy and biopsy specimens of human skin were fixed overnight in alcoholic Bouin''s solution, embedded in paraffin, cut at 7 [mu] deparaffinized, hydrated to 70% alcohol, and treated as follows[long dash]stained 2 hours in a mixture consisting of: 0.2% orcein in 70% alcohol and 1% HC1 (cone.), 125 ml; 5% hematoxylin in absolute alcohol, 40 ml; 6% FeClg in water, 25 ml; and aqueous I2-KI (1:2:100), 25 ml[long dash]rinsed in distilled water until the excess stain was removed[long dash]differentiated in 1.2% FeC13, 5-15 sec-washed in running water, 5 min[long dash]differentiation completed in 0.01% HC1 acid-alcohol, 1 min-a dip in 95% alcohol-distilled water, 2 min[long dash]0.25% aqueous metanil yellow, 5[long dash]10 sec[long dash]a 95% alcohol dip[long dash]dehydrated in absolute alcohol, xylene, and mounted in a resinous medium. The technic combines the orcein of Pinkus'' stain and the hematoxylin mixture of Verhoeff into a single staining solution and gives sharp and reliable results for both coarse and extremely delicate elastic fibers. These stain purple; nuclei, violet; and background, yellow. The stain allows the use of formalin, Bouin''s fluid and Zenker-formol fixation. The results have been consistent in other primates as well as in man.