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. 2024:2813:167-188.
doi: 10.1007/978-1-0716-3890-3_12.

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

Chiara Toniolo  1 , Daniel Sage  2 , John D McKinney  1 , Neeraj Dhar  3   4   5
Affiliations

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

Chiara Toniolo et al. Methods Mol Biol. 2024.

Abstract

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.

Keywords: Host-pathogen interaction; Image analysis; Infection; Live imaging; Macrophages; Time-lapse microscopy; Mycobacterium tuberculosis.

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References

    1. Bandekar AC, Subedi S, Ioerger TR et al (2020) Cell-cycle-associated expression patterns predict gene function in mycobacteria. Curr Biol 30:3961–3971 - DOI - PubMed - PMC
    1. Naveen G, Peerapur BV (2012) Comparison of the Lowenstein-Jensen Medium, the Middlebrook 7H10 Medium and MB/BacT for the isolation of Mycobacterium Tuberculosis (MTB) from clinical specimens. J Clin Diagn Res 6:1704–1709 - PubMed - PMC
    1. VanderVen BC, Fahey RJ, Lee W et al (2015) Novel inhibitors of cholesterol degradation in mycobacterium tuberculosis reveal how the bacterium’s metabolism is constrained by the intracellular environment. PLoS Pathog. https://doi.org/10.1371/journal.ppat.1004679
    1. Koul A, Vranckx L, Dhar N et al (2014) Delayed bactericidal response of Mycobacterium tuberculosis to bedaquiline involves remodelling of bacterial metabolism. Nat Commun. https://doi.org/10.1038/ncomms4369
    1. Andréanne L, Anthony V, Shi-Yan FC et al (2018) Optimized background regimen for treatment of active tuberculosis with the next-generation Benzothiazinone Macozinone (PBTZ169). Antimicrob Agents Chemother 62:e00840–e00818

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