Tracking single particles and elongated filaments with nanometer precision.
Biophys J
; 100(11): 2820-8, 2011 Jun 08.
Article
em En
| MEDLINE
| ID: mdl-21641328
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 â¼2 nm when determining the filament centerline and â¼9 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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Cinesinas
/
Proteínas de Drosophila
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Nanotecnologia
/
Microtúbulos
Limite:
Animals
Idioma:
En
Revista:
Biophys J
Ano de publicação:
2011
Tipo de documento:
Article