Single Molecule Fluorescence in Membrane Biology.
Our Chapter in Single Molecule Biology by Alex Knight.
Single molecule fluorescence (SMF) methods have made major contributions to the current understanding of membrane biology. This chapter discusses findings obtained by SMF into the mechanisms that govern membrane structure, cell signaling, ion channel gating, and vesicle transport. It also considers how recent advances in single molecule imaging technology could be applied to membranes and used to identify technological limitations currently inhibiting the field. Several key characteristics of membranes and their components make them well suited to single molecule approaches, including their accessibility to in vivo studies due to their location at the cell surface, the availability of model cell membranes for in vitro studies, and the confinement of molecules to 2D motion. SMF imaging of membranes in vivo requires the specific attachment of fluorescent labels to proteins or lipids of interest within the membrane of live cells, and the primary challenges facing membrane studies are related to labeling and detection technology.