The visualization of fluorescent proteins in living cells by video intensification microscopy (VIM)

Abstract

A highly sensitive television camera (silicon intensifier target) has been combined with fluorescence microscopy to examine living cultured cells. This system is termed Video Intensification Microscopy (VIM). By using very small amounts of excitation light, one limits the damage to living cells from excessive illumination and is able to visualize fluorescence probes for periods up to 24 hr without bleaching. With VIM, the cellular uptake and fate of two rhodamine-labeled proteins, concanavalin A and alpha2 macroglobulin, have been followed for up to 24 hr. These proteins were first located in endocytic vesicles with a low phase density. Later, at 24 hr, alpha2 macroglobulin was located in phase-dense structures, probably secondary lysosomes. Both the fluorescent endocytic vesicles and lysosomes were observed to undergo saltatory motion. VIM combined with fluorescence promises to have a widespread application in the study of the behavior of living cells.