Quantification of membrane trafficking on a 3D cytoskeleton network in living cells

Recent progress in biology and microscopy has made possible to acquire multidimensional data on rapid cellular activities. Unfortunately, data analysis, ultimate step to describe the observed biological process, still remains a major bottleneck. We here present a novel method devoted to quantitatively investigate the membrane trafficking, by monitoring vesicular structures moving along a three dimensional cytoskeleton network. It is based on kymogram analysis in order to extract the consistent part of the temporal information from the initial data. It is able to handle typical complex situations, like the case of a dense set of vesicles moving at various velocities, fusing and dissociating together or with other cell compartments. Application of this method to the Rab6 protein, a GTPase involved in the regulation of intracellular membrane trafficking, gives a large number of cellular dynamic information