FRAP to Characterize Molecular Diffusion and Interaction in Various Membrane Environments.

ABSTRACT: Fluorescence recovery after photobleaching (FRAP) is a standard method used to study the dynamics of lipids and proteins in artificial and cellular membrane systems. The advent of confocal microscopy two decades ago has made quantitative FRAP easily available to most laboratories. Usually, a single bleaching pattern/area is used and the corresponding recovery time is assumed to directly provide a diffusion coefficient, although this is only true in the case of unrestricted Brownian motion. Here, we propose some general guidelines to perform FRAP experiments under a confocal microscope with different bleaching patterns and area, allowing the experimentalist to establish whether the molecules undergo Brownian motion (free diffusion) or whether they have restricted or directed movements. Usin

SUBMITTER: Pincet F 

PROVIDER: S-EPMC4936743 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature