AUTHOR=Iino Ryota , Matsumoto Yoshimi , Nishino Kunihiko , Yamaguchi Akihito , Noji Hiroyuki 

TITLE=Design of a large-scale femtoliter droplet array for single-cell analysis of drug-tolerant and drug-resistant bacteria

JOURNAL=Frontiers in Microbiology

VOLUME=4

YEAR=2013

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2013.00300

DOI=10.3389/fmicb.2013.00300

ISSN=1664-302X

ABSTRACT=<p>Single-cell analysis is a powerful method to assess the heterogeneity among individual cells, enabling the identification of very rare cells with properties that differ from those of the majority. In this Methods Article, we describe the use of a large-scale femtoliter droplet array to enclose, isolate, and analyze individual bacterial cells. As a first example, we describe the single-cell detection of drug-tolerant persisters of <italic>Pseudomonas aeruginosa</italic> treated with the antibiotic carbenicillin. As a second example, this method was applied to the single-cell evaluation of drug efflux activity, which causes acquired antibiotic resistance of bacteria. The activity of the MexAB-OprM multidrug efflux pump system from <italic>Pseudomonas aeruginosa</italic> was expressed in <italic>Escherichia coli</italic> and the effect of an inhibitor D13-9001 were assessed at the single cell level.</p>