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- W1966793007 abstract "Understanding the dynamics of intracellular vesicles is essential for studies of cellular internalization, intracellular transport, and the movement of proteins between organelles. Of particular interest are the dynamics that result in the fusion of vesicles as extracellular cargo is transported through the endocytic pathway. We use two-color single particle tracking fluorescence microscopy to study populations of vesicles that exist as both isolated vesicles and fused hybrid vesicles. Specifically, we are interested in the relationship between Rab7-vesicles, LAMP1-vesicles, and Rab7-LAMP1-hybrid vesicles. The majority of these vesicles exist as hybrid vesicles, which limits the ability of static measurements and standard biochemical approaches to resolve differences between these three populations. We propose that while these vesicle populations are highly colocalized, they can be distinguished by their dynamics during the short periods when they are not colocalized. To this end, individual vesicles, labeled with distinct variants of GFP, were tracked and analyzed. An algorithm was developed to obtain the total and linear distance traveled by the vesicle, travel direction relative to its eventual fusion site, and efficiency in reaching its destination. Significant differences in these measurements suggest LAMP1-vesicles fuse more efficiently than Rab7-vesicles. This approach will prove useful in future studies for resolving the differences in highly colocalized populations of vesicles." @default.
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- W1966793007 date "2011-02-01" @default.
- W1966793007 modified "2023-09-26" @default.
- W1966793007 title "Dynamic Behavior of Intracellular Vesicles Probed with Two-Color Single Particle Tracking" @default.
- W1966793007 doi "https://doi.org/10.1016/j.bpj.2010.12.3463" @default.
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