State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, College of Chemistry, Nankai University, Tianjin, 300071, China.
Analyst. 2019 Jan 28;144(3):859-871. doi: 10.1039/c8an01420e.
Generally, the message elucidated by the conventional analytical methods overlooks the heterogeneity of single objects, where the behavior of individual molecules is shielded. With the advent of optical microscopy imaging techniques, it is possible to identify, visualize and track individual molecules or nanoparticles under a biological environment with high temporal and spatial resolution. In this work, we summarize the commonly adopted optical microscopy techniques for bio-analytical assays in living cells, including total internal reflection fluorescence microscopy (TIRFM), super-resolution optical microscopy (SRM), and dark-field optical microscopy (DFM). The basic principles of these methods and some recent interesting applications in molecular detection and single-particle tracking are introduced. Moreover, the development in high-dimensional optical microscopy to achieve three-dimensional (3-D) as well as sub-diffraction localization and tracking of biomolecules is also highlighted.
一般来说，传统分析方法所阐明的信息忽略了单个物体的异质性，其中个体分子的行为被掩盖了。随着光学显微镜成像技术的出现，有可能在具有高时空分辨率的生物环境中识别、可视化和跟踪单个分子或纳米颗粒。在这项工作中，我们总结了在活细胞中进行生物分析的常用光学显微镜技术，包括全内反射荧光显微镜（TIRFM）、超分辨率光学显微镜（SRM）和暗场光学显微镜（DFM）。介绍了这些方法的基本原理以及在分子检测和单粒子跟踪方面的一些最新有趣应用。此外，还强调了多维光学显微镜在实现三维（3-D）以及生物分子的亚衍射定位和跟踪方面的发展。
