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A Robust Method for Generating, Quantifying, and Testing Large Numbers of Escherichia coli Persisters
Silke R Vedelaar1, Jakub L Radzikowski1,2, Matthias Heinemann3
1Molecular Systems Biology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands.
Methods in Molecular Biology (Clifton, N.J.)
|September 30, 2021
Summary
Researchers developed a method to generate large populations of antibiotic-tolerant bacterial persister cells using nutrient shifts. This breakthrough facilitates the study of persister cells, crucial for understanding antibiotic tolerance.
Area of Science:
- Microbiology
- Bacterial Physiology
- Antibiotic Resistance
Background:
- Bacteria can develop antibiotic tolerance, a critical challenge in medicine.
- Persister cells, a small subpopulation exhibiting tolerance, are difficult to study due to their scarcity.
Purpose of the Study:
- To present a protocol for generating high-frequency populations of antibiotic-tolerant persister cells.
- To introduce methods for quantifying persister cell fractions and assessing antibiotic tolerance.
Main Methods:
- Utilizing abrupt nutrient shifts to induce persister cell formation in Escherichia coli.
- Employing population-averaging experimental techniques and flow cytometry for analysis.
Main Results:
- Successfully generated Escherichia coli populations with a high proportion of antibiotic-tolerant cells.
- Established methods to quantify the percentage of cells entering the tolerant state and assess tolerance levels.
Conclusions:
- The nutrient-switch approach effectively creates homogeneous persister cell populations.
- Developed methodologies will advance research into bacterial persister cell phenotypes and antibiotic tolerance mechanisms.
