Skip to Main content Skip to Navigation
Book sections

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.
Complete list of metadatas

Cited literature [11 references]  Display  Hide  Download

https://hal.inrae.fr/hal-02918510
Contributor : Arnaud Chastanet <>
Submitted on : Thursday, August 20, 2020 - 4:03:40 PM
Last modification on : Friday, October 23, 2020 - 4:47:04 PM

File

21-Cornilleau-Methods (Ch 8)-2...
Files produced by the author(s)

Identifiers

Collections

Citation

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⟩

Share

Metrics

Record views

85

Files downloads

20