Nature Methods
- 5, 687 - 694 (2008)
Published online: 6 July 2008; | doi:10.1038/nmeth.1233
Dynamic multiple-target tracing to probe spatiotemporal cartography of cell membranesArnauld Sergé1, 2, 3, Nicolas Bertaux4, 5, 6, Hervé Rigneault4, 5 & Didier Marguet1, 2, 31
Centre d'Immunologie de Marseille-Luminy, Université de la Méditerranée, UMR 6102, Parc scientifique de Luminy, Case 906, F-13288 Marseille cedex 09, France. 2
Institut National de la Santé et de la Recherche Médicale, Unité Mixte de Recherche 631, UMR 6102, Parc scientifique de Luminy, Case 906, F-13288 Marseille cedex 09, France. 3
Centre National de la Recherche Scientifique, UMR 6102, Parc scientifique de Luminy, Case 906, F-13288 Marseille cedex 09, France. 4
Institut Fresnel, Université Paul Cézanne, UMR 6133, Domaine Universitaire de Saint Jérôme, F-13397 Marseille cedex 20, France. 5
Centre National de la Recherche Scientifique, UMR 6133, Domaine Universitaire de Saint Jérôme, F-13397 Marseille cedex 20, France. 6
École Centrale Marseille, Technopôle de Château-Gombert, 38, rue Frédéric Joliot Curie, F-13451 Marseille cedex 20, France.
Correspondence should be addressed to Didier Marguet marguet@ciml.univ-mrs.fr Although the highly dynamic and mosaic organization of the plasma membrane is well-recognized, depicting a resolved, global view of this organization remains challenging. We present an analytical single-particle tracking (SPT) method and tool, multiple-target tracing (MTT), that takes advantage of the high spatial resolution provided by single-fluorophore sensitivity. MTT can be used to generate dynamic maps at high densities of tracked particles, thereby providing global representation of molecular dynamics in cell membranes. Deflation by subtracting detected peaks allows detection of lower-intensity peaks. We exhaustively detected particles using MTT, with performance reaching theoretical limits, and then reconnected trajectories integrating the statistical information from past trajectories. We demonstrate the potential of this method by applying it to the epidermal growth factor receptor (EGFR) labeled with quantum dots (Qdots), in the plasma membrane of live cells. We anticipate the use of MTT to explore molecular dynamics and interactions at the cell membrane.
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