High-Throughput Single-Cell Analysis of Vaccinia Virus Infection

Liam M Howell; Timothy P Newsome

doi:10.1007/978-1-0716-4160-6_15

High-Throughput Single-Cell Analysis of Vaccinia Virus Infection

Methods Mol Biol. 2025:2860:229-240. doi: 10.1007/978-1-0716-4160-6_15.

Authors

Liam M Howell¹, Timothy P Newsome^{2

3}

Affiliations

¹ School of Life and Environmental Science, The University of Sydney, Camperdown, NSW, Australia.
² School of Life and Environmental Science, The University of Sydney, Camperdown, NSW, Australia. [email protected].
³ Sydney Institute for Infectious Diseases, The University of Sydney, Camperdown, NSW, Australia. [email protected].

PMID: 39621271
DOI: 10.1007/978-1-0716-4160-6_15

Abstract

Here, we describe a robust and flexible method for analyzing the infection of single cells with a fluorescent-reporter virus, with real-time tracking of infected cells through microscopy. We subsequently generate quantitative data from the resulting time-lapse microscopy, and harness these data to generate biologically meaningful parameters at scale. Our methodology offers a practical solution for researchers seeking to understand the complexities and variability of virus-host interactions at a granular level.

Keywords: Fluorescence microscopy; Image analysis; Infection dynamics; Poxvirus; Single cell; Single-cell isolation; Time series data modeling; Vaccinia virus.

MeSH terms

High-Throughput Screening Assays / methods
Host-Pathogen Interactions
Humans
Microscopy, Fluorescence / methods
Single-Cell Analysis* / methods
Time-Lapse Imaging / methods
Vaccinia / virology
Vaccinia virus* / physiology