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Single-Cell Real-Time Visualization and Quantification of Perylene Bioaccumulation in Microorganisms
Xin Jin1, Xuejun Guo1, Deshu Xu1
1State Key Laboratory of Environment Simulation, School of Environment, Beijing Normal University , No. 19 Xinjiekouwai Street, Beijing 100875, China.
Environmental Science & Technology
|May 19, 2017
Summary
Single-molecule fluorescence microscopy revealed distinct perylene bioaccumulation patterns in bacteria. Live Escherichia coli showed high cell-to-cell variation due to efflux systems, unlike Staphylococcus aureus.
Area of Science:
- Microbiology
- Biochemistry
- Biophysics
Background:
- Perylene is a polycyclic aromatic hydrocarbon with potential environmental and biological impacts.
- Understanding its bioaccumulation in bacteria is crucial for environmental risk assessment.
- Bacterial responses to xenobiotics can vary significantly between species and even individual cells.
Purpose of the Study:
- To visualize and quantify real-time perylene bioaccumulation in single bacterial cells.
- To investigate cell-to-cell heterogeneity in perylene accumulation dynamics.
- To elucidate the mechanisms underlying differential perylene bioaccumulation in Escherichia coli and Staphylococcus aureus.
Main Methods:
- Single-molecule fluorescence microscopy (SMFM) was employed for high-sensitivity, high-temporal-resolution imaging.
- A microfluidic flow chamber with precise temperature control was utilized.
- Live and dead bacterial cells (E. coli and S. aureus) were analyzed to assess the role of cell viability.
Main Results:
- Escherichia coli exhibited significant cell-to-cell heterogeneity in perylene accumulation (C.V = 1.40), observed only in live cells.
- Staphylococcus aureus showed low heterogeneity in perylene accumulation, regardless of cell viability (C.V = 0.36).
- Tol C-associated efflux systems in E. coli were identified as key factors for lower accumulation and high heterogeneity.
Conclusions:
- Bacterial bioaccumulation of perylene is highly heterogeneous in live E. coli, driven by active efflux mechanisms.
- S. aureus demonstrates passive diffusion of perylene due to a lack of efficient efflux systems.
- SMFM is a powerful tool for studying dynamic molecular processes and heterogeneity in microbial systems.
