Widom Julia R, Dhakal Soma, Heinicke Laurie A, Walter Nils G
Single Molecule Analysis Group, Department of Chemistry, University of Michigan, 930 N. University Ave., Ann Arbor, MI, 48109-1055, USA.
Arch Toxicol. 2014 Nov;88(11):1965-85. doi: 10.1007/s00204-014-1357-9. Epub 2014 Sep 12.
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.
毒理学是研究化学物质对生物体不利影响的高度跨学科领域。它需要灵敏的工具来检测此类影响。单分子荧光检测工具在20世纪90年代首次应用后，很快就因其对许多不同科学探究领域有巨大贡献的潜力而得到认可。在此期间，该领域的技术进步带来了不断提高的空间和时间分辨率，并使单分子荧光能够应用于越来越复杂的系统，如活细胞。在这篇综述中，我们概述了实现最常见版本的单分子荧光检测所需的光学组件。然后，我们讨论了其在酶学与结构研究、系统生物学和纳米技术方面的当前应用，介绍了每个研究领域独特的技术考量以及近期值得关注的成果。我们还强调了未来的发展方向，这些方向有可能通过进一步利用单分子荧光显微镜的能力来彻底改变这些研究领域。
