Rapid visualization of Acanthamoeba using fluorescein-conjugated lectins

J. B. Robin, R. Chan and B. R. Andersen
Department of Ophthalmology, University of Illinois College of Medicine at Chicago.

We investigated the efficacy of fluorescein-conjugated lectins (FCLs) for the rapid visualization of Acanthamoeba species. Cultures of Acanthamoeba castellani, Acanthamoeba culbertsoni, and Acanthamoeba polyphaga were established on nonnutrient agar plates supplemented with Escherichia coli. Maximal trophozoite populations were established four to five days after initial subculturing; mature cysts were routinely noted three to six days later. At various time points, trophozoites and/or cysts were harvested and suspended in Page's saline. Following a previously described protocol, samples of the suspensions were placed on glass slides and incubated with one of a panel of 14 FCLs. At the end of the incubation period, the slides were examined using an epifluorescence microscope. Concanavalin A brightly stained both cysts and trophozoites. Wheat germ agglutinin brightly stained cysts but only minimally stained trophozoites. These results suggest that FCLs may be useful for rapid visualization of Acanthamoeba organisms in corneal infections.