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A robust method for generating, quantifying, and testing large numbers of escherichia coli persisters.

Title: A robust method for generating, quantifying, and testing large numbers of escherichia coli persisters.
Authors: Vedelaar, SR
Radzikowski, JL
Heinemann, M
Item Type: Chapter
Abstract: Bacteria can exhibit phenotypes that render them tolerant against antibiotics. However, often only a few cells of a bacterial population show the so-called persister phenotype, which makes it difficult to study this health-threatening phenotype. We recently found that certain abrupt nutrient shifts generate Escherichia coli populations that consist almost entirely of antibiotic-tolerant cells. These nearly homogeneous persister cell populations enable assessment with population-averaging experimental methods, such as high-throughput methods. In this chapter, we provide a detailed protocol for generating a large fraction of tolerant cells using the nutrient-switch approach. Furthermore, we describe how to determine the fraction of cells that enter the tolerant state upon a sudden nutrient shift and we provide a new way to assess antibiotic tolerance using flow cytometry. We envision that these methods will facilitate research into the important and exciting phenotype of bacterial persister cells.
Issue Date: 30-Sep-2021
URI: http://hdl.handle.net/10044/1/92346
DOI: 10.1007/978-1-0716-1621-5_3
Start Page: 41
End Page: 62
Journal / Book Title: Methods in Molecular Biology
Volume: 2357
Copyright Statement: © The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2021
Keywords: Antibiotic tolerance
Escherichia coli
Flow cytometry
Nutrient shift
Persister cells
Tolerant cells
Antibiotic tolerance
Escherichia coli
Flow cytometry
Nutrient shift
Persister cells
Tolerant cells
Developmental Biology
0399 Other Chemical Sciences
0601 Biochemistry and Cell Biology
Publication Status: Published
Open Access location: https://link.springer.com/protocol/10.1007%2F978-1-0716-1621-5_3
Online Publication Date: 2021-09-30
Appears in Collections:Department of Surgery and Cancer