Abstract
Tracking the diffusive mobility of individual membrane proteins within the cell offers a route to unravel how protein function is sculpted by the architecture of the membrane and its constituents. To meet this challenge fully, however, demands a high resolution in both space and time which exceeds the capabilities of fluorescence microscopies. Interferometric scattering (iSCAT) microscopy is an emerging fluorescence-free technique highly suited to the sensitive detection and precise tracking of nanoscopic biological entities, including viruses, vesicles, and individual proteins, both in artificial and live-cell contexts. In this presentation, we demonstrate successful implementation of iSCAT microscopy to track proteins within the plasma membrane of the living cell to nanometric precision in all three-dimensions whilst at fast microsecond speeds. We will discuss our recent results resolving nanoscopic membrane confinement, uptake and prolonged directed diffusion along a filopodium for the epidermal growth factor receptor protein. In addition, we will also discuss the future prospects and challenges for iSCAT microscopy in exploring single-molecule dynamics within living cells.
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