Analyzing persister physiology with fluorescence-activated cell sorting

Mehmet A. Orman, Theresa C. Henry, Christina J. Decoste, Mark P. Brynildsen

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Scopus citations

Abstract

Bacterial persisters are phenotypic variants that exhibit an impressive ability to tolerate antibiotics. Persisters are hypothesized to cause relapse infections, and therefore, understanding their physiology may lead to novel therapeutics to treat recalcitrant infections. However, persisters have yet to be isolated due to their low abundance, transient nature, and similarity to the more highly abundant viable but non-culturable cells (VBNCs), resulting in limited knowledge of their phenotypic state. This technical hurdle has been addressed through the use of fluorescence-activated cell sorting (FACS) and quantification of persister levels in the resulting sorted fractions. These assays provide persister phenotype distributions, which can be compared to the phenotype distributions of the entire population, and can also be used to examine persister heterogeneity. Here, we describe two detailed protocols for analysis of persister physiology with FACS. One protocol assays the metabolic state of persisters using a fluorescent metabolic stain, whereas the other assays the growth state of persisters with use of a fluorescent protein.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages83-100
Number of pages18
DOIs
StatePublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1333
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Keywords

  • Antibiotic
  • Fluorescence-activated cell sorting (FACS)
  • Persister
  • Phenotypic heterogeneity
  • Redox sensor green (RSG)
  • Viable but non-culturable cell (VBNC)

Fingerprint Dive into the research topics of 'Analyzing persister physiology with fluorescence-activated cell sorting'. Together they form a unique fingerprint.

  • Cite this

    Orman, M. A., Henry, T. C., Decoste, C. J., & Brynildsen, M. P. (2016). Analyzing persister physiology with fluorescence-activated cell sorting. In Methods in Molecular Biology (pp. 83-100). (Methods in Molecular Biology; Vol. 1333). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-2854-5_8