Development of Ultrafast Camera-based Single-Molecule Imaging Unravels Plasma Membrane Structure and Function in Live Cells

Abstract

Regulation of molecular diffusion in the plasma membrane (PM) is fundamentally important for enabling many PM functions. An ultrafast camera system was developed for single fluorescent-molecule imaging at time resolutions better than 0.1 ms, which confirmed the actin-based PM compartmentalization. The camera system was applied to develop ultrafast live-cell PALM and dSTORM. They revealed the mesoscopic dynamic organization of the focal adhesion (FA), in which the FA has an archipelago architecture of ≈30-nm diameter FA-protein islands loosely clustered into ≈300-nm diameter functional units in the compartmentalized fluid membrane (74 nm vs 110 nm outside the FA).