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Issue 11, 2014
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Dielectrophoresis-based purification of antibiotic-treated bacterial subpopulations

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Abstract

Persistence of bacteria during antibiotic therapy is a widespread phenomenon, of particular importance in refractory mycobacterial infections such as leprosy and tuberculosis. Persistence is characterized by the phenotypic tolerance of a subpopulation of bacterial cells to antibiotics. Characterization of these “persister” cells is often difficult due to the transient, non-heritable nature of the phenotype and due to the presence of contaminating material from non-persisting cells, which usually comprise the larger fraction. In this study, we use 3D carbon-electrode arrays for dielectrophoresis-based separation of intact cells from damaged cells, revealed by differential staining with propidium iodide, and we use this procedure to purify intact cells from cultures of Mycobacterium smegmatis treated with isoniazid, a frontline anti-tuberculosis drug. The method presented in this study could be used for rapid label-free enrichment of intact persister cells from antibiotic-treated cultures while preserving the metastable persister phenotype. This approach would facilitate the downstream analysis of low-frequency subpopulations of cells using conventional omics techniques, such as transcriptomic and proteomic analysis.

Graphical abstract: Dielectrophoresis-based purification of antibiotic-treated bacterial subpopulations

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Supplementary files

Article information


Submitted
24 Jan 2014
Accepted
09 Apr 2014
First published
10 Apr 2014

Lab Chip, 2014,14, 1850-1857
Article type
Paper
Author version available

Dielectrophoresis-based purification of antibiotic-treated bacterial subpopulations

M. Elitas, R. Martinez-Duarte, N. Dhar, J. D. McKinney and P. Renaud, Lab Chip, 2014, 14, 1850
DOI: 10.1039/C4LC00109E

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