Visualization and quantification of vesicle trafficking on a three‐dimensional cytoskeleton network in living cells
- 16 March 2007
- journal article
- Published by Wiley in Journal of Microscopy
- Vol. 225 (3), 214-228
- https://doi.org/10.1111/j.1365-2818.2007.01723.x
Abstract
Recent progress in biology and microscopy has made it possible to acquire multidimensional data on rapid cellular activities. Unfortunately, the data analysis needed to describe the observed biological process still remains a major bottleneck. We here present a novel method of studying membrane trafficking by monitoring vesicular structures moving along a three-dimensional cytoskeleton network. It allows the dynamics of such structures to be qualitatively and quantitatively investigated. Our tracking method uses kymogram analysis to extract the consistent part of the temporal information and to allow the meaningful representation of vesicle dynamics. A fully automatic extension of this method, together with a statistical tool for dynamic comparisons, allows the precise analysis and comparison of overall speed distributions and directions. It can handle typical complex situations, such as a dense set of vesicles moving at various velocities, fusing and dissociating with each other or with other cell compartments. The overall approach has been characterized and validated on synthetic data. We chose the Rab6A protein, a GTPase involved in the regulation of intracellular membrane trafficking, as a molecular model. The application of our method to GFP-Rab6A stable cells acquired using fast four-dimensional deconvolution video-microscopy gives considerable cellular dynamic information unreachable using other techniques.Keywords
This publication has 31 references indexed in Scilit:
- Dynamics and Spatial Organization of Endosomes in Mammalian CellsPhysical Review Letters, 2005
- Bicaudal-D regulates COPI-independent Golgi–ER transport by recruiting the dynein–dynactin motor complexNature Cell Biology, 2002
- Converging Populations of F-Actin Promote Breakage of Associated Microtubules to Spatially Regulate Microtubule Turnover in Migrating CellsCurrent Biology, 2002
- Dual-wavelength fluorescent speckle microscopy reveals coupling of microtubule and actin movements in migrating cellsThe Journal of cell biology, 2002
- How Rab proteins link motors to membranesNature Cell Biology, 2002
- The Human Kinesin-Like Protein RB6K Is under Tight Cell Cycle Control and Is Essential for CytokinesisMolecular and Cellular Biology, 2001
- Rab6 Coordinates a Novel Golgi to ER Retrograde Transport Pathway in Live CellsThe Journal of cell biology, 1999
- An unbiased detector of curvilinear structuresIEEE Transactions on Pattern Analysis and Machine Intelligence, 1998
- Sonar tracking of multiple targets using joint probabilistic data associationIEEE Journal of Oceanic Engineering, 1983
- An algorithm for tracking multiple targetsIEEE Transactions on Automatic Control, 1979