Quantification of Microbial Fluorescent Sensors During Live Intracellular Infections

Erez Mills*, Erik Petersen

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

Abstract

The interaction of pathogens with their eukaryotic hosts during intracellular growth is critical to many diseases. However, the relative scarcity of pathogen biomolecules versus the abundant host biomolecule concentration can make quantitative evaluation of pathogen intracellular responses difficult. Recent years have seen an explosion in utilization of fluorescent proteins to serve as transcriptional reporters and biosensors for quantification of pathogen responses. Here, we describe a method to establish a fluorescent assay quantifying pathogen behavior during intracellular infection and to quantify these results at a single cell level. The sensitivity of these fluorescent assays permits the live observation of changing pathogen responses, while the ability to measure at a single cell level uncovers subpopulations of pathogens whose existence may be missed during the population-level assays often required to accumulate sufficient pathogen biomolecules for analysis.

Original languageAmerican English
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages119-131
Number of pages13
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2427
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Bibliographical note

Funding Information:
Research in the Mills lab is supported by grants from the Israel Science Foundation (grant number 1272/20), the Israeli Ministry of Science and Technology (grant number 88638), and the US-Israel Binational Agricultural Research and Development Fund (grant number IS-5242-20). Research in the Petersen lab is supported by the US-Israel Binational Agricultural Research and Development Fund (grant number IS-5242-20). EM is chair of the Vigevani Senior Lectureship in Animal Sciences.

Publisher Copyright:
© 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

  • Cyclic-di-GMP
  • Fluorescent reporter
  • Intracellular pathogen
  • Live cell microscopy
  • Salmonella
  • Single-cell quantification

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