Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA.
Department of Chemical Engineering, Myongji University, Yongin 17058, Republic of Korea.
Sensors (Basel). 2023 Sep 6;23(18):7691. doi: 10.3390/s23187691.
Single-molecule imaging technologies, especially those based on fluorescence, have been developed to probe both the equilibrium and dynamic properties of biomolecules at the single-molecular and quantitative levels. In this review, we provide an overview of the state-of-the-art advancements in single-molecule fluorescence imaging techniques. We systematically explore the advanced implementations of in vitro single-molecule imaging techniques using total internal reflection fluorescence (TIRF) microscopy, which is widely accessible. This includes discussions on sample preparation, passivation techniques, data collection and analysis, and biological applications. Furthermore, we delve into the compatibility of microfluidic technology for single-molecule fluorescence imaging, highlighting its potential benefits and challenges. Finally, we summarize the current challenges and prospects of fluorescence-based single-molecule imaging techniques, paving the way for further advancements in this rapidly evolving field.
单分子成像技术，特别是基于荧光的单分子成像技术，已经被开发出来以在单分子和定量水平上探测生物分子的平衡和动态特性。在这篇综述中，我们提供了单分子荧光成像技术的最新进展概述。我们系统地探索了使用广泛应用的全内反射荧光（TIRF）显微镜的体外单分子荧光成像技术的高级实现。这包括对样品制备、钝化技术、数据收集和分析以及生物应用的讨论。此外，我们深入研究了微流控技术与单分子荧光成像的兼容性，强调了其潜在的优势和挑战。最后，我们总结了基于荧光的单分子成像技术目前面临的挑战和前景，为这个快速发展的领域的进一步发展铺平了道路。
