Application Of Digitized Fluorescence Microscopy And Video Photobleaching To Study Membrane Dynamics During Cell Locomotion

Our laboratory is concerned with understanding the dynamic structure of the plasma membrane with particular reference to the movement of membrane constituents during cell locomotion. We employ digitized fluorescence microscopy (DFM) alone or in combination with fluorescence recovery after photobleaching (FRAP) to investigate individual cells. DFM is really a new form of light microscopy in that the distribution of individual classes of ions, molecules, and macromolecules can be followed in single, living cells. By employing fluorescent antibodies to define antigens or fluorescent analogs of cellular constituents as well as ultra-sensitive, electronic image detectors and video image averaging to improve signal to noise, fluorescent images of living cells can be acquired over an extended period without significant fading and loss of cell viability. FRAP allows the measurement of translational mobility of membrane and cytoplasmic molecules in small regions of single, living cells.