Single-Cell Detection and Collection of Persister Bacteria in a Directly Accessible Femtoliter Droplet Array

Ryota Iino; Shouichi Sakakihara; Yoshimi Matsumoto; Kunihiko Nishino

doi:10.1007/978-1-4939-2854-5_9

Single-Cell Detection and Collection of Persister Bacteria in a Directly Accessible Femtoliter Droplet Array

Methods Mol Biol. 2016:1333:101-9. doi: 10.1007/978-1-4939-2854-5_9.

Authors

Ryota Iino^{1

2}, Shouichi Sakakihara³, Yoshimi Matsumoto⁴, Kunihiko Nishino⁴

Affiliations

¹ Okazaki Institute for Integrative Bioscience, Institute for Molecular Science, National Institutes of Natural Sciences, 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 Laboratory of Microbiology and Infectious Diseases, Institute of Scientific and Industrial Research, Osaka University, Osaka, Japan.

PMID: 26468103
DOI: 10.1007/978-1-4939-2854-5_9

Abstract

A directly accessible femtoliter droplet array as a platform for single-cell detection and collection of persister bacteria is described. Device microfabrication, femtoliter droplet array formation and concomitant enclosure of single cells, long-term culture and observation of single cells in droplets, and collection of identified persisters from single droplets are described in detail.

Keywords: Drug tolerance; Microdevice; Microdroplet array; Microfabrication; Micropipette; Optical microscopy; Persister; Pseudomonas aeruginosa; Single-cell analysis.

MeSH terms

Anti-Bacterial Agents / pharmacology*
Drug Tolerance / genetics*
Humans
Microbial Sensitivity Tests
Microtechnology / methods*
Pseudomonas aeruginosa / drug effects
Pseudomonas aeruginosa / growth & development
Single-Cell Analysis / methods*

Substances

Anti-Bacterial Agents