title: Tracking single particles and elongated filaments with nanometer precision.
abstract: 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 ∼2 nm when determining the filament centerline and ∼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.
authors.affiliation: B CUBE-Center of Innovation Competence, Technische Universität Dresden, Dresden, Germany.
authors.forename: Felix
authors.initials: F
authors.lastname: Ruhnow
authors.forename: David
authors.initials: D
authors.lastname: Zwicker
authors.forename: Stefan
authors.initials: S
authors.lastname: Diez
mesh_terms: D000465:Algorithms; D000818:Animals; D029721:Drosophila Proteins / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D016547:Kinesins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D009068:Movement; D036103:Nanotechnology / Q000379:methods*; D012984:Software
chemical_list: D029721:Drosophila Proteins; D005456:Fluorescent Dyes; D016547:Kinesins