Detection of individual fluorescently labeled reovirions in living cells

Reovirus serotype 1 (Lang) can be conjugated with rhodamine B or fluorescein isothiocyanate in a way that preserves viral infectivity. We have used epifluorescence microscopy to detect individual virions bound to the surface of cells and to follow in real time the early stages of reovirus infection in living cells. Following uptake of the virus into endocytic vesicles, the movement of these vesicles can be observed readily. The vesicle movement is inhibited by nocodazole or colchicine, consistent with previous findings that the movement of intracellular vesicles is often microtubule-based.