Monitoring and quantifying dynamic physiological processes in live neurons using fluorescence recovery after photobleaching

Authors

  • Kevin Staras,

    1. School of Life Sciences, University of Sussex, Brighton, UK
    Search for more papers by this author
  • Dan Mikulincer,

    1. Department of Physiology and Cell Biology, Faculty of Health Sciences, Zlotowski Center for Neuroscience, Ben-Gurion University of the Negev, Beer-Sheva, Israel
    Search for more papers by this author
  • Daniel Gitler

    Corresponding author
    1. Department of Physiology and Cell Biology, Faculty of Health Sciences, Zlotowski Center for Neuroscience, Ben-Gurion University of the Negev, Beer-Sheva, Israel
    • School of Life Sciences, University of Sussex, Brighton, UK
    Search for more papers by this author

Address correspondence and reprint requests to Daniel Gitler, Department of Physiology and Cell Biology, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel. E-mail: gitler@bgu.ac.il

Abstract

The direct visualization of subcellular dynamic processes is often hampered by limitations in the resolving power achievable with conventional microscopy techniques. Fluorescence recovery after photobleaching has emerged as a highly informative approach to address this challenge, permitting the quantitative measurement of the movement of small organelles and proteins in living functioning cells, and offering detailed insights into fundamental cellular phenomena of physiological importance. In recent years, its implementation has benefited from the increasing availability of confocal microscopy systems and of powerful labeling techniques based on genetically encoded fluorescent proteins or other chemical markers. In this review, we present fluorescence recovery after photobleaching and related techniques in the context of contemporary neurobiological research and discuss quantitative and semi-quantitative approaches to their interpretation.

Ancillary