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Single-molecule tools for enzymology, structural biology, systems biology and nanotechnology: an update

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Abstract

Toxicology is the highly interdisciplinary field studying the adverse effects of chemicals on living organisms. It requires sensitive tools to detect such effects. After their initial implementation during the 1990s, single-molecule fluorescence detection tools were quickly recognized for their potential to contribute greatly to many different areas of scientific inquiry. In the intervening time, technical advances in the field have generated ever-improving spatial and temporal resolution and have enabled the application of single-molecule fluorescence to increasingly complex systems, such as live cells. In this review, we give an overview of the optical components necessary to implement the most common versions of single-molecule fluorescence detection. We then discuss current applications to enzymology and structural studies, systems biology, and nanotechnology, presenting the technical considerations that are unique to each area of study, along with noteworthy recent results. We also highlight future directions that have the potential to revolutionize these areas of study by further exploiting the capabilities of single-molecule fluorescence microscopy.

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Acknowledgments

This work was supported in part by National Institutes of Health Grants R01 GM062357, R01 GM098023 and R21 AI109791, and by the Department of Defense MURI Award W911NF-12-1-0420 to N.G.W.

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The authors declare that they have no conflict of interest.

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  1. Single Molecule Analysis Group, Department of Chemistry, University of Michigan, 930 N. University Ave., Ann Arbor, MI, 48109-1055, USA

    Julia R. Widom, Soma Dhakal, Laurie A. Heinicke & Nils G. Walter

  2. Single Molecule Analysis in Real-Time (SMART) Center, University of Michigan, 930 N. University Ave., Ann Arbor, MI, 48109-1055, USA

    Nils G. Walter

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  1. Julia R. Widom
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Widom, J.R., Dhakal, S., Heinicke, L.A. et al. Single-molecule tools for enzymology, structural biology, systems biology and nanotechnology: an update. Arch Toxicol 88, 1965–1985 (2014). https://doi.org/10.1007/s00204-014-1357-9

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