Quantification of Mycobacterium tuberculosis Growth in Cell-Based Infection Assays by Time-Lapse Fluorescence Microscopy

C. Toniolo, D. Sage, J.D. McKinney, N. Dhar

Methods in Molecular Biology, vol. 2813, Walker J.M., Ed., Springer, ch. 12, pp. 167–188, 2024.

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Quantification of Mycobacterium tuberculosis (Mtb) growth dynamics in cell-based in vitro infection models is traditionally carried out by measurement of colony forming units (CFU). However, Mtb being an extremely slow growing organism (16–24 h doubling time), this approach requires at least 3 weeks of incubation to obtain measurable readouts. In this chapter, we describe an alternative approach based on time-lapse microscopy and quantitative image analysis that allows faster quantification of Mtb growth dynamics in host cells. In addition, this approach provides the capability to capture other readouts from the same experimental setup, such as host cell viability, bacterial localization as well as the dynamics of propagation of infection between the host cells.

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AUTHOR="Toniolo, C. and Sage, D. and McKinney, J.D. and Dhar, N.",
TITLE="Quantification of {\textit{Mycobacterium tuberculosis}} Growth in
	Cell-Based Infection Assays by Time-Lapse Fluorescence Microscopy",
BOOKTITLE="Methods in Molecular Biology",
PUBLISHER="Springer",
YEAR="2024",
editor="Walker, J.M.",
volume="2813",
series="Thakur, A. (Ed.), Intracellular Pathogens",
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chapter="12",
pages="167--188",
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DOI: 10.1007/978-1-0716-3890-3_12