Processing TIRF Microscopy Images to Characterize the Dynamics and Morphology of Bacterial Actin-Like Assemblies

Authors :: Cyrille Billaudeau
Arnaud Chastanet
Rut Carballido-López
MICrobiologie de l'ALImentation au Service de la Santé (MICALIS)
AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
Source :: Cytoskeleton Dynamics-Methods and Protocols, Methods in Molecular Biology, Methods in Molecular Biology-Cytoskeleton Dynamics, Cytoskeleton dynamics: methods and protocols, methods in molecular biology, Cytoskeleton dynamics: methods and protocols, methods in molecular biology, pp.135-145, 2020, ⟨10.1007/978-1-0716-0219-5_9⟩, Methods in Molecular Biology ISBN: 9781071602188
Publication Year :: 2020
Abstract: Total internal reflection fluorescence (TIRF) microscopy allows the visualization of the dynamic membrane-associated actin-like MreB filaments in live bacterial cells with high temporal resolution. This chapter describes computerized analysis methods to quantitatively characterize the dynamics and morphological properties of MreB assemblies. These include how to (1) segment bacterial cells, (2) perform single-particle tracking (SPT) of MreB filamentous structures, (3) classify their dynamic modes using mean squared displacement (MSD) analysis, and (4) measure their dimensions and orientation.

Full Text Access

Tools