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Recent advances in experimental design and data analysis to characterize prokaryotic motility
Bacterial motility plays a key role in important cell processes such as chemotaxis and biofilm formation, but is challenging to quantify due to the small size of the individual microorganisms and the complex interplay of biological and physical factors that influence motility phenotypes. Swimming, t...
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Published in: | Journal of microbiological methods 2023-01, Vol.204, p.106658-106658, Article 106658 |
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Main Authors: | , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | Bacterial motility plays a key role in important cell processes such as chemotaxis and biofilm formation, but is challenging to quantify due to the small size of the individual microorganisms and the complex interplay of biological and physical factors that influence motility phenotypes. Swimming, the first type of motility described in bacteria, still remains largely unquantified. Light microscopy has enabled qualitative characterization of swimming patterns seen in different strains, such as run and tumble, run-reverse-flick, run and slow, stop and coil, and push and pull, which has allowed for elucidation of the underlying physics. However, quantifying these behaviors (e.g., identifying run distances and speeds, turn angles and behavior by surfaces or cell-cell interactions) remains a challenging task. A qualitative and quantitative understanding of bacterial motility is needed to bridge the gap between experimentation, omics analysis, and bacterial motility theory. In this review, we discuss the strengths and limitations of how phase contrast microscopy, fluorescence microscopy, and digital holographic microscopy have been used to quantify bacterial motility. Approaches to automated software analysis, including cell recognition, tracking, and track analysis, are also discussed with a view to providing a guide for experimenters to setting up the appropriate imaging and analysis system for their needs.
•Tracking of actively swimming microorganisms, especially in 3D, remains a challenging problem.•Design of a successful tracking experiment requires careful consideration of hardware, camera, and processing software.•Extremophilic microorganisms may require special considerations for temperature control and labeling.•Detection of microorganisms in video images and linking of tracks are separate considerations.•This article summarizes currently available hardware and software to serve as a guide to designing a tracking experiment. |
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ISSN: | 0167-7012 1872-8359 |
DOI: | 10.1016/j.mimet.2022.106658 |