A single-cell drug efflux assay in bacteria by using a directly accessible femtoliter droplet array

Active efflux of drugs, such as antibiotics, from a cell is one of the major mechanisms that cause multi-drug resistance in bacteria. Here we report a method to assess drug efflux activity in individual Escherichia coli cells enclosed and isolated in a directly accessible femtoliter droplet array wi...

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Bibliographic Details
Published in:Lab on a chip 2012-10, Vol.12 (20), p.3923-3929
Main Authors: Iino, Ryota, Hayama, Kohei, Amezawa, Hiromi, Sakakihara, Shouichi, Kim, Soo Hyeon, Matsumono, Yoshimi, Nishino, Kunihiko, Yamaguchi, Akihito, Noji, Hiroyuki
Format: Article
Language:English
Subjects:
Accessibility
Anti-Bacterial Agents - pharmacology
Arrays
Bacteria
Biological Assay - instrumentation
Biological Assay - methods
Droplets
Drug Resistance, Multiple, Bacterial - physiology
Drugs
Efflux
Escherichia
Escherichia coli - genetics
Escherichia coli - metabolism
Fluorescent Dyes - pharmacology
Genes
Microbial Sensitivity Tests - instrumentation
Microbial Sensitivity Tests - methods
Microfluidic Analytical Techniques - instrumentation
Microfluidic Analytical Techniques - methods
Pumps
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Summary:Active efflux of drugs, such as antibiotics, from a cell is one of the major mechanisms that cause multi-drug resistance in bacteria. Here we report a method to assess drug efflux activity in individual Escherichia coli cells enclosed and isolated in a directly accessible femtoliter droplet array with a fluorogenic compound. The inhibitory effect of a chemical compound on an exogenously expressed efflux pump system from pathogenic bacteria was easily detected at the single-cell level. We also present a proof-of-principle experiment to screen for the gene encoding a drug efflux pump by collecting individual droplets containing single cells in which the drug efflux activity was restored after introduction of the exogenous gene from pathogenic bacteria. Our approach will be a useful tool to screen novel pump inhibitors and efflux pump genes, and to overcome infectious diseases caused by multi-drug-resistant bacteria.
ISSN:1473-0197
1473-0189
DOI:10.1039/c2lc40394c