Journal
METHODS
Volume 94, Issue -, Pages 75-84Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2015.09.002
Keywords
Podosomes; Macrophages; Multiparticle tracking; Force; Mechanosensing; Collective dynamics
Funding
- Fondation pour la Recherche Medicale [FRM DEQ-2011-0421312]
- l'Agence Nationale de la Recherche [ANR 2010-01301, ANR14-CE11-0020-02]
- European Community [HEALTH-F4-2011-282095-Tarkinaid]
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Podosomes are submicron adhesive and mechanosensitive structures formed by macrophages, dendritic cells and osteoclasts that are capable of protruding into the extracellular environment. Built of an F-actin core surrounded by an adhesion ring, podosomes assemble in a network interconnected by acto-myosin cables. They have been shown to display spatiotemporal instability as well as protrusion force oscillations. To analyse the entire population of these unstable structures, we have designed an automated multi-particle tracking adapted to both topographical and fluorescence data. Here we describe in detail this approach and report the measurements of individual and collective characteristics of podosome ensembles, providing an integrated picture of their activity from the complementary angles of organisation, dynamics, mobility and mechanics. We believe that this will lead to a comprehensive view of podosome collective behaviour and deepen our knowledge about the significance of mechanosensing mediated by protrusive structures. (C) 2015 Elsevier Inc. All rights reserved.
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