title: Actin filament dynamics using microfluidics.
abstract: We describe how combining microfluidics with TIRF and epifluorescence microscopy can greatly facilitate the quantitative analysis of actin assembly dynamics and its regulation, as well as the exploration of issues that were often out of reach with standard single-filament microscopy, such as the kinetics of processes linked to actin self-assembly or the kinetics of interaction with regulators. We also show how the viscous drag force exerted by fluid flowing on the filaments can be calibrated in order to assess the mechanosensitivity of end-binding protein machineries such as formins or adhesion proteins. We also discuss how microfluidics, in conjunction with other techniques, could be used to address the mechanism of coordination between heterogeneous populations of filaments, or the behavior of individual filaments during regulated treadmilling. These techniques also can be applied to study the assembly and regulation of other cytoskeletal polymers such as microtubules, septins, intermediate filaments, as well as the transport of cargoes by molecular motors under a flow-produced load.
authors.affiliation: Laboratoire d'Enzymologie et Biochimie Structurales, CNRS, Gif-sur-Yvette, France. Electronic address: carlier@lebs.cnrs-gif.fr.
authors.forename: Marie-France
authors.initials: MF
authors.lastname: Carlier
authors.affiliation: Laboratoire d'Enzymologie et Biochimie Structurales, CNRS, Gif-sur-Yvette, France. Electronic address: romet@lebs.cnrs-gif.fr.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Romet-Lemonne
authors.affiliation: Laboratoire d'Enzymologie et Biochimie Structurales, CNRS, Gif-sur-Yvette, France. Electronic address: jegou@lebs.cnrs-gif.fr.
authors.forename: Antoine
authors.initials: A
authors.lastname: Jégou
mesh_terms: D008841:Actin Cytoskeleton / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D004867:Equipment Design; D007700:Kinetics; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D013499:Surface Properties
keywords: Actin networks; Actin regulatory proteins; Force measurements; Microfluidics; Single actin filament assembly dynamics; Surface passivation and functionalization; TIRF microscopy