Zirconyl Hematoxylin: a New Stain for Acidic Mucins

Abstract

Most stains for acidic mucins are time-consuming to prepare and have poor stability. Zirconyl hematoxylin is easily prepared and works for a year or more. It is made by adding 5 ml freshly-made 0.1% aqueous sodium iodate, 400 mg zirconyl chloride octahydrate, and 40 ml 25% aqueous glycerol, in that order, to 100 mg of hematoxylin in 5 ml of absolute ethanol and stirring for 5 min. Stain 10 min and do not "blue" the stain. Chlorazole black or kernechtrot and fast green are good counterstains. Zirconyl hematoxylin stains acidic mucins violet or red violet, regardless of how they are fixed. It stains the same mucins as alcian blue in mouse and sheep salivary glands. It shows goblet cells in mouse rectum as well as alcian blue. It stains the same stomach regions in a lizard as alcian blue. Like alcian blue and colloidal iron, zirconyl hematoxylin stains the mucin of cancerous prostate, but not normal prostate.

Use of an improved zirconyl hematoxylin stain in the diagnosis of Barrett's esophagus

“…The original zirconyl hematoxylin used the oxidant to hematoxylin ratio recommended by Lillie [10,23] which left most of the hematoxylin intact for later oxidation by atmospheric oxygen during the working life of the solution. It worked well on the acidic mucins of rodent salivary glands [21] and the abnormal acidic mucin of prostatic cancer [22].…”

“…It worked well on the acidic mucins of rodent salivary glands [21] and the abnormal acidic mucin of prostatic cancer [22].…”

“…Zirconyl hematoxylin also stains acidic mucins [21,22]. While the staining intensity of the original formulation is adequate for aiding in the diagnosis of adenocarcinoma of the prostate [22], it is too pale for rapid screening of esophageal biopsies.…”