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.
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Acknowledgment
This work was supported by a consolidator grant from the European Research Council (ERC COG) under the European Union’s Horizon 2020 research and innovation program (grant agreement No. 772178) to R.C.-L.
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Billaudeau, C., Chastanet, A., Carballido-López, R. (2020). Processing TIRF Microscopy Images to Characterize the Dynamics and Morphology of Bacterial Actin-Like Assemblies. In: Maiato, H. (eds) Cytoskeleton Dynamics. Methods in Molecular Biology, vol 2101. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0219-5_9
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DOI: https://doi.org/10.1007/978-1-0716-0219-5_9
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