Large-Scale Femtoliter Droplet Array for Single Cell Efflux Assay of Bacteria

Ryota Iino; Shouichi Sakakihara; Yoshimi Matsumoto; Kunihiko Nishino

doi:10.1007/978-1-4939-7454-2_18

Large-Scale Femtoliter Droplet Array for Single Cell Efflux Assay of Bacteria

Methods Mol Biol. 2018:1700:331-341. doi: 10.1007/978-1-4939-7454-2_18.

Authors

Ryota Iino^{1

2}, Shouichi Sakakihara³, Yoshimi Matsumoto⁴, Kunihiko Nishino^{5

6}

Affiliations

¹ Okazaki Institute for Integrative Bioscience, Institute for Molecular Science, National Institutes of Natural Sciences, Okazaki, Aichi, 444-8787, Japan. iino@ims.ac.jp.
² Department of Functional Molecular Science, School of Physical Sciences, The Graduate University for Advanced Studies (SOKENDAI), Kanagawa, 240-0193, Japan. iino@ims.ac.jp.
³ Technical Division, Institute of Scientific and Industrial Research, Osaka University, Osaka, Japan.
⁴ Department of Biomolecular Science and Regulation, Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Osaka, Japan.
⁵ Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka, 567-0047, Japan.
⁶ Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, 565-0871, Japan.

PMID: 29177839
DOI: 10.1007/978-1-4939-7454-2_18

Abstract

Large-scale femtoliter droplet array as a platform for single cell efflux assay of bacteria is described. Device microfabrication, femtoliter droplet array formation and concomitant enclosure of single bacterial cells, fluorescence-based detection of efflux activity at the single cell level, and collection of single cells from droplet and subsequent gene analysis are described in detail.

Keywords: AcrAB-TolC; Drug efflux; Escherichia coli; Fluorescence; MexAB-OprM; Microdroplet array; Microfabrication; Optical microscopy; Pseudomonas aeruginosa; Salmonella enterica; Single cell analysis.

MeSH terms

Bacterial Proteins / analysis
Escherichia coli / metabolism
Membrane Transport Proteins / analysis*
Microbial Sensitivity Tests
Pseudomonas aeruginosa / metabolism
Salmonella enterica / metabolism
Single-Cell Analysis / instrumentation*
Single-Cell Analysis / methods
Tissue Array Analysis / instrumentation*
Tissue Array Analysis / methods

Substances

Bacterial Proteins
Membrane Transport Proteins