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Population and Single-Cell Analysis of Antibiotic Persistence in Escherichia coli
Published on: March 24, 2023
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Precision single cell analysis to characterize host dependent antimicrobial response heterogeneity in physiological
Ryuichiro Abe1, Jyong-Huei Lee2, Siew Mei Chin2
1Department of Emergency Medicine, Stanford University School of Medicine, Stanford, CA, USA. syang5@stanford.edu.
Lab on a Chip
|February 3, 2025
Summary
Antimicrobial susceptibility testing (AST) using a novel microfluidic platform reveals significant heterogeneity in bacterial responses to antibiotics. This single-cell approach enables personalized antimicrobial treatment strategies for multidrug-resistant pathogens.
Area of Science:
- Microbiology
- Biotechnology
- Infectious Diseases
Background:
- Antimicrobial stewardship is crucial for managing multidrug-resistant pathogens.
- Current phenotypic antimicrobial susceptibility testing (AST) methods have limitations in mimicking physiological conditions and resolving population heterogeneity.
- There is a need for advanced AST techniques to guide personalized antimicrobial therapy.
Purpose of the Study:
- To investigate the heterogeneous responses of uropathogenic bacteria to antimicrobials in a host-dependent manner.
- To develop and validate a microfluidic platform for single-cell analysis of bacterial responses to antimicrobial treatment.
- To assess the potential for personalized antimicrobial treatment based on single-cell kinetics.
Main Methods:
- Development of a microfluidic gel encapsulation platform for single-cell imaging.
- Culturing uropathogenic bacteria in individual urine samples under antimicrobial treatment.
- Real-time monitoring of bacterial growth and killing kinetics at the single-cell level.
- Medium exchange experiments to evaluate the recovery of surviving bacterial subpopulations.
Main Results:
- Demonstrated substantial heterogeneity in bacterial time-kill kinetics in response to antimicrobials, influenced by host factors.
- Successfully resolved heterogeneous subpopulations with distinct responses to antimicrobial pressure using the microfluidic platform.
- Observed varying abilities of surviving cells to resume growth after antimicrobial removal, highlighting population heterogeneity.
Conclusions:
- The microfluidic platform provides a powerful tool for high-resolution, single-cell analysis of antimicrobial responses.
- Bacterial populations exhibit significant heterogeneity in their response to antimicrobials, necessitating a move beyond bulk measurements.
- This approach lays the groundwork for precision medicine in antimicrobial treatment, enabling personalized therapeutic strategies.
