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Environ Sci Technol. 2017 Jun 6;51(11):6211-6219. doi: 10.1021/acs.est.7b02070. Epub 2017 May 26.

Single-Cell Real-Time Visualization and Quantification of Perylene Bioaccumulation in Microorganisms.

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State Key Laboratory of Environment Simulation, School of Environment, Beijing Normal University , No. 19 Xinjiekouwai Street, Beijing 100875, China.
Biodynamic Optical Imaging Center, School of Life Sciences, Peking University , No. 5 Yiheyuan Road, Beijing 100871, China.


Bioaccumulation of perylene in Escherichia coli and Staphylococcus aureus was visualized and quantified in real time with high sensitivity at high temporal resolution. For the first time, single-molecule fluorescence microscopy (SMFM) with a microfluidic flow chamber and temperature control has enabled us to record the dynamic process of perylene bioaccumulation in single bacterial cells and examine the cell-to-cell heterogeneity. Although with identical genomes, individual E. coli cells exhibited a high degree of heterogeneity in perylene accumulation dynamics, as shown by the high coefficient of variation (C.V = 1.40). This remarkable heterogeneity was exhibited only in live E. coli cells. However, the bioaccumulation of perylene in live and dead S. aureus cells showed similar patterns with a low degree of heterogeneity (C.V = 0.36). We found that the efflux systems associated with Tol C played an essential role in perylene bioaccumulation in E. coli, which caused a significantly lower accumulation and a high cell-to-cell heterogeneity. In comparison with E. coli, the Gram-positive bacteria S. aureus lacked an efficient efflux system against perylene. Therefore, perylene bioaccumulation in S. aureus was simply a passive diffusion process across the cell membrane.

[Indexed for MEDLINE]

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