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

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Book cover Mycobacteria Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2314))

Abstract

Studies on cell-to-cell phenotypic variation in microbial populations, with individuals sharing the same genetic background, provide insights not only on bacterial behavior but also on the adaptive spectrum of the population. Phenotypic variation is an innate property of microbial populations, and this can be further amplified under stressful conditions, providing a fitness advantage. Furthermore, phenotypic variation may also precede a latter step of genetic-based diversification, resulting in the transmission of the most beneficial phenotype to the progeny. While population-wide studies provide a measure of the collective average behavior, single-cell studies, which have expanded over the last decade, delve into the behavior of smaller subpopulations that would otherwise remain concealed. In this chapter, we describe approaches to carry out spatiotemporal analysis of individual mycobacterial cells using time-lapse microscopy. Our method encompasses the fabrication of a microfluidic device; the assembly of a microfluidic system suitable for long-term imaging of mycobacteria; and the quantitative analysis of single-cell behavior under varying growth conditions. Phenotypic variation is conceivably associated to the resilience and endurance of mycobacterial cells. Therefore, shedding light on the dynamics of this phenomenon, on the transience or stability of the given phenotype, on its molecular bases and its functional consequences, offers new scope for intervention.

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Acknowledgments

This work was supported by the Institut Pasteur and by ANR grants (ANR-10-LABX-62-IBEID and ANR-17-CE11-0007-01) to GM. ND acknowledges support from the Swiss South African Joint Research Program of the Swiss National Science Foundation (Project IZLSZ3_170912). GM & ND were supported by the Innovative Medicines Initiative 2 Joint Undertaking (JU) under grant agreement No 853989. The JU receives support from the European Union’s Horizon 2020 research and innovation programme and EFPIA and Global Alliance for TB Drug Development non profit organization, Bill & Melinda Gates Foundation, University of Dundee.

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Authors and Affiliations

  1. Microbial Individuality and Infection Group, Cell Biology and Infection Department, Microbiology Department, Institut Pasteur, Paris, France

    Giulia Manina

  2. School of Life Sciences, Swiss Federal Institute of Technology in Lausanne (EPFL), Lausanne, Switzerland

    Neeraj Dhar

Authors
  1. Giulia Manina
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  2. Neeraj Dhar
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Corresponding author

Correspondence to Giulia Manina .

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Editors and Affiliations

  1. Center for Global Infectious Disease Research, Seattle Children’s Research Institute, Seattle, WA, USA

    Tanya Parish

  2. Center for Global Infectious Disease Research, Seattle Children’s Research Institute, Seattle, WA, USA

    Anuradha Kumar

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Manina, G., Dhar, N. (2021). Single-Cell Analysis of Mycobacteria Using Microfluidics and Time-Lapse Microscopy. In: Parish, T., Kumar, A. (eds) Mycobacteria Protocols. Methods in Molecular Biology, vol 2314. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1460-0_8

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  • DOI: https://doi.org/10.1007/978-1-0716-1460-0_8

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1459-4

  • Online ISBN: 978-1-0716-1460-0

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