The Combined Gram-Pappenheim Stain as Modified for Films and Paraffin Sections

Abstract

A combination of the Gram-Pappenheim stains for the examination of gonorrheal pus, cellular exudate and paraffin sections of formalin-fixed tissues has been described elsewhere (Scudder and Lisa, 1931). The crystal violet solution was made stable for the first time by employing phosphate buffers on the acid side of neutrality, and a stable counterstain was prepared for the first time from National Aniline dyes, ethylated methyl green and pyronin yellowish. Original findings were demonstrated by means of color plate I and II (Scudder, 1931) to show gonococci, pneumococci and cells in smears, and formalin-fixed tissue brought down to water in the usual way. A new color plate is published herewith to show the microscopic appearance of cells, Gram-positive and Gram-negative bacteria, higher bacteria, fungi and spermatozoa in the study of genitourinary and gynecological cases. The method has a value in the field of medical jurisprudence. Crystals were well demonstrated, especially those resulting from sulfa drug therapy. The National Aniline methyl green batches numbered NG 10, 11, 13 to 19, and their batches of pyronin numbered NP 5 to 10 were found consistently stable. Earlier dyes were found either too purple or too blue for the technic and the most satisfactory dyes were found to require a ripening time of several days and could be prepared in amounts of from 1 to 4 liters and stored indefinitely without preservatives.