A Triple Stain for Deoxyribonucleic Acid, Polysaccharides and Proteins

Tissues from mammalian, amphibian, insect and plant sources were fixed in formalin or in Carnoy's, Helly's, or Smith's fluid and processed in the usual manner to mounted paraffin sections. These were stained by the sequence: Feulgen reaction (using azure-A-Schiff reagent); periodic acid Schiff (with basic-fuchsin-Schiff reagent); and a 0.02% solution of naphthol yellow S in 1 % acetic acid. Nuclei stained blue to green; polysaccharides, red; and proteins, yellow. Thus, deoxyribonucleic acid, vicinal hydroxy and hydroxy-amino groups, and free basic groups of proteins were demonstrated selectively. Of the tissues tested, stomach, intestine, kidney, thyroid gland and plant root tips were most strikingly stained.