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Article
Nature Methods - 5, 695 - 702 (2008)
Published online: 20 July 2008; | doi:10.1038/nmeth.1237

Robust single-particle tracking in live-cell time-lapse sequences

Khuloud Jaqaman1, Dinah Loerke1, Marcel Mettlen1, Hirotaka Kuwata2, Sergio Grinstein2, Sandra L Schmid1 & Gaudenz Danuser1

1  Department of Cell Biology, The Scripps Research Institute, 10550 N. Torrey Pines Rd, La Jolla, California 92037, USA.

2  Department of Cell Biology, The Hospital for Sick Children, 555 University Ave., Toronto M5G1X8, Canada.

Correspondence should be addressed to Khuloud Jaqaman kjaqaman@scripps.edu

Single-particle tracking (SPT) is often the rate-limiting step in live-cell imaging studies of subcellular dynamics. Here we present a tracking algorithm that addresses the principal challenges of SPT, namely high particle density, particle motion heterogeneity, temporary particle disappearance, and particle merging and splitting. The algorithm first links particles between consecutive frames and then links the resulting track segments into complete trajectories. Both steps are formulated as global combinatorial optimization problems whose solution identifies the overall most likely set of particle trajectories throughout a movie. Using this approach, we show that the GTPase dynamin differentially affects the kinetics of long- and short-lived endocytic structures and that the motion of CD36 receptors along cytoskeleton-mediated linear tracks increases their aggregation probability. Both applications indicate the requirement for robust and complete tracking of dense particle fields to dissect the mechanisms of receptor organization at the level of the plasma membrane.

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Axiovert 200 inverted epifluorescence microscope (Zeiss)
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Nature Methods
ISSN: 1548-7091
EISSN: 1548-7105
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