Journal
BIOPHYSICAL JOURNAL
Volume 100, Issue 11, Pages 2820-2828Publisher
CELL PRESS
DOI: 10.1016/j.bpj.2011.04.023
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Funding
- European Research Council [242933]
- European Commission [NMP4-SL-2009-228971]
- Deutsche Forschungsgemeinschaft [DI 1226/3]
- Max-Planck-Society
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Recent developments in image processing have greatly advanced our understanding of biomolecular processes in vitro and in vivo. In particular, using Gaussian models to fit the intensity profiles of nanometer-sized objects have enabled their two-dimensional localization with a precision in the one-nanometer range. Here, we present an algorithm to precisely localize curved filaments whose structures are characterized by subresolution diameters and micrometer lengths. Using surface-immobilized microtubules, fluorescently labeled with rhodamine, we demonstrate positional precisions of similar to 2 nm when determining the filament centerline and similar to 9 nm when localizing the filament tips. Combined with state-of-the-art single particle tracking we apply the algorithm 1), to motor-proteins stepping on immobilized microtubules, 2), to depolymerizing microtubules, and 3), to microtubules gliding over motor-coated surfaces.
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