Chapter 11
Flow Cytometry Analysis of Mycobacteria
and Mycobacteria-Infected Immune Cells
Sydney L. Solomon and Bryan D. Bryson
Abstract
Flow cytometry enables the measurement of tens of features on individual cells from complex mixtures.
Flow cytometry enables high-throughput quantification of cell size, gene and protein expression. In the
case of studies of host-pathogen interactions, this tool provides a facile way of identifying cells that have
been successfully infected by a pathogen. Several recent technological advances have greatly improved
throughput and the number of features that can be simultaneously monitored by this technique. Here, we
describe common workflows to study Mycobacterium tuberculosis heterogeneity and host-M. tuberculosis
interactions using flow cytometry and related technologies.
Key words Flow cytometry, Mycobacteria, Mass cytometry, Image cytometry, Immunology
1 Introduction
Flow cytometry involves the measurement of biochemical and
physical characteristics of a population of cells or particles with
single-cell resolution [1]. Several disciplines in biology have benefited tremendously from the ability to acquire tens of measurements
per cell from millions of cells as well as isolate specific populations of
interest via sorting. Traditionally, in flow cytometry and similar
technologies, a cell suspension is delivered via a fluidics system to
the cytometer. The cytometer then measures the light scattering
properties and additional information pertaining to the intensity of
specific fluorochromes (flow cytometry) or mass tags (mass cytometry) [2, 3].
In the study of mycobacteria and the immune response to
mycobacteria, flow cytometry and related techniques have become
increasingly common [4–6]. Flow cytometry can be utilized to
monitor biophysical properties of individual mycobacteria and
identify infected host cells. Furthermore, uninfected and infected
cells can be probed for host and mycobacterial gene and protein
expression dynamics concurrently, as well as biomolecules such as
Tanya Parish and Anuradha Kumar (eds.), Mycobacteria Protocols, Methods in Molecular Biology, vol. 2314,
https://doi.org/10.1007/978-1-0716-1460-0_11, © Springer Science+Business Media, LLC, part of Springer Nature 2021
261
Flow Cytometry Analysis of Mycobacteria
and Mycobacteria-Infected Immune Cells
Sydney L. Solomon and Bryan D. Bryson
Abstract
Flow cytometry enables the measurement of tens of features on individual cells from complex mixtures.
Flow cytometry enables high-throughput quantification of cell size, gene and protein expression. In the
case of studies of host-pathogen interactions, this tool provides a facile way of identifying cells that have
been successfully infected by a pathogen. Several recent technological advances have greatly improved
throughput and the number of features that can be simultaneously monitored by this technique. Here, we
describe common workflows to study Mycobacterium tuberculosis heterogeneity and host-M. tuberculosis
interactions using flow cytometry and related technologies.
Key words Flow cytometry, Mycobacteria, Mass cytometry, Image cytometry, Immunology
1 Introduction
Flow cytometry involves the measurement of biochemical and
physical characteristics of a population of cells or particles with
single-cell resolution [1]. Several disciplines in biology have benefited tremendously from the ability to acquire tens of measurements
per cell from millions of cells as well as isolate specific populations of
interest via sorting. Traditionally, in flow cytometry and similar
technologies, a cell suspension is delivered via a fluidics system to
the cytometer. The cytometer then measures the light scattering
properties and additional information pertaining to the intensity of
specific fluorochromes (flow cytometry) or mass tags (mass cytometry) [2, 3].
In the study of mycobacteria and the immune response to
mycobacteria, flow cytometry and related techniques have become
increasingly common [4–6]. Flow cytometry can be utilized to
monitor biophysical properties of individual mycobacteria and
identify infected host cells. Furthermore, uninfected and infected
cells can be probed for host and mycobacterial gene and protein
expression dynamics concurrently, as well as biomolecules such as
Tanya Parish and Anuradha Kumar (eds.), Mycobacteria Protocols, Methods in Molecular Biology, vol. 2314,
https://doi.org/10.1007/978-1-0716-1460-0_11, © Springer Science+Business Media, LLC, part of Springer Nature 2021
261