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Chapitre d'ouvrage

Methods for Studying Membrane-Associated Bacterial Cytoskeleton Proteins In Vivo by TIRF Microscopy

Abstract : MreB proteins are actin homologs present in nonspherical bacteria. They assemble into membrane-associated discrete filamentous structures that exhibit different dynamic behaviors along the bacterial sidewalls. Total internal reflection fluorescence (TIRF) microscopy, a sensitive method for studying molecular events at cell surfaces with high contrast and temporal resolution, is a method of choice to characterize the localization and dynamics of cortical MreB assemblies in vivo. This chapter describes the methods for visualizing fluorescently tagged MreB proteins in live Bacillus subtilis cells. We detail how to (1) grow B. subtilis strains for reproducible TIRF observations, (2) immobilize cells on agarose pads and (3) in CellASIC ® microfluidic plates, and (4) acquire TIRF images and time lapses.
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https://hal.inrae.fr/hal-02918510
Déposant : Arnaud Chastanet <>
Soumis le : jeudi 20 août 2020 - 16:03:40
Dernière modification le : vendredi 23 octobre 2020 - 16:47:04

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Charlène Cornilleau, Arnaud Chastanet, Cyrille Billaudeau, Rut Carballido-Lopez. Methods for Studying Membrane-Associated Bacterial Cytoskeleton Proteins In Vivo by TIRF Microscopy. Cytoskeleton Dynamics: Methods and Protocols, Methods in Molecular Biology, pp.123-133, 2020, ⟨10.1007/978-1-0716-0219-5_8⟩. ⟨hal-02918510⟩

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