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Single-Cell Analysis of Mycobacteria Using Microfluidics and Time-Lapse Microscopy

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Mycobacteria Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1285))

Abstract

The crucial role of phenotypic heterogeneity in bacterial physiology and adaptive responses has required the introduction of new ways to investigate bacterial individuality. Time-lapse microscopy is a powerful technique for evaluating phenotypic diversity in bacteria at the single-cell level, whether exploring the dynamics of gene expression and protein localization or characterizing the heterogeneous phenotypic response to perturbations. Here, we present protocols to carry out time-lapse imaging of mycobacteria at the single-cell level using either agarose pads or customized microfluidic devices. The sequences of images obtained can be analyzed using programs such as ImageJ and allow the investigator not only to extract various parameters of growth and gene expression dynamics but also to unravel the physiological basis behind phenomenon such as persistence against stresses.

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Author information

Authors and Affiliations

  1. Laboratory of Microbiology and Microsystems, School of Life Sciences, Swiss Federal Institute of Technology in Lausanne (EPFL), Station 19, Office SV 3832, Lausanne, CH-1015, Switzerland

    Neeraj Dhar & Giulia Manina

Authors
  1. Neeraj Dhar
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  2. Giulia Manina
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Corresponding author

Correspondence to Neeraj Dhar .

Editor information

Editors and Affiliations

  1. Infectious Disease Research Inst (IDRI), Seattle, Washington, USA

    Tanya Parish

  2. Infectious Disease Research Inst (IDRI), Seattle, Washington, USA

    David M. Roberts

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Dhar, N., Manina, G. (2015). Single-Cell Analysis of Mycobacteria Using Microfluidics and Time-Lapse Microscopy. In: Parish, T., Roberts, D. (eds) Mycobacteria Protocols. Methods in Molecular Biology, vol 1285. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2450-9_14

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  • DOI: https://doi.org/10.1007/978-1-4939-2450-9_14

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2449-3

  • Online ISBN: 978-1-4939-2450-9

  • eBook Packages: Springer Protocols

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