Foot's Silver and Acridine-Orange Fluorescence Staining of Pneumocystis Carinii in Smears and in Sections

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Abstract
Pneumocystis carinii was stained intensely by the modification of Hortega's silver carbonate method for reticulum. Paraffin sections and smears were fixed in formalin, then processed through all the steps of the staining technique as given in McClung's Handbook (Jones 1950), p. 258. Fluorescence microscopy was also very helpful in disclosing the parasite in air-dried smears. These, were processed through descending grades of ethanol and then stained in 0.01% Acridine orange in phosphate buffer, pH 6.0, for 2 min. The microorganism appeared in a brownish to red range of fluorescence, whereas disintegrated cellular nuclei ranged from green to yellow. A standard binocular microscope equipped with a high-pressure 200-watt mercury vapor burner (HBO 200, Osram) and the necessary filters (BG 12 as exciting and OG 5 or OG 4 as barrier filters) were used.

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