Department of Physics, Biological Physics Research Group, University of Oxford, Oxford, UK.
Phys Biol. 2010 Aug 4;7(3):031001. doi: 10.1088/1478-3975/7/3/031001.
Single-molecule fluorescence microscopy is currently one of the most popular methods in the single-molecule toolbox. In this review, we discuss recent advances in fluorescence instrumentation and assays: these methods are characterized by a substantial increase in complexity of the instrumentation or biological samples involved. Specifically, we describe new multi-laser and multi-colour fluorescence spectroscopy and imaging techniques, super-resolution microscopy imaging and the development of instruments that combine fluorescence detection with other single-molecule methods such as force spectroscopy. We also highlight two pivotal developments in basic and applied biosciences: the new information available from detection of single molecules in single biological cells and exciting developments in fluorescence-based single-molecule DNA sequencing.
单分子荧光显微镜是目前单分子工具包中最受欢迎的方法之一。在这篇综述中，我们讨论了荧光仪器和测定法的最新进展：这些方法的特点是仪器或涉及的生物样本的复杂性有了实质性的提高。具体来说，我们描述了新的多激光和多色荧光光谱和成像技术、超分辨率显微镜成像以及将荧光检测与其他单分子方法（如力谱学）相结合的仪器的发展。我们还强调了基础和应用生物科学中的两个关键进展：从单个生物细胞中单分子检测中获得的新信息，以及基于荧光的单分子 DNA 测序的令人兴奋的发展。
