Peng Sijia, Wang Wenjuan, Chen Chunlai
School of Life Sciences, Tsinghua-Peking Joint Center for Life Sciences, and Beijing Advanced Innovation Center for Structural Biology, Tsinghua University, Beijing, 100084, P.R. China.
School of Life Sciences and Technology Center for Protein Sciences, Tsinghua University, Beijing, 100084, P.R. China.
Chemistry. 2018 Jan 24;24(5):1002-1009. doi: 10.1002/chem.201704065. Epub 2017 Nov 30.
Fluorescence-based single-molecule techniques have become widely used tools to reveal dynamic processes of biomolecules and elucidate their molecular mechanisms. However, the concentration upper limit of labeled species that can be used in single-molecule fluorescence measurements is at the low nm range, which is below the Michaelis constants of many enzymatic reactions and physiological concentrations of many biomolecules. Such discrepancy limits the application of single-molecule fluorescence tools. Several techniques have been developed to break the concentration barrier. In this Concept, we focus on reviewing fundamental principles of these techniques and wish to inspire development of new and better tools to achieve this goal.
基于荧光的单分子技术已成为揭示生物分子动态过程并阐明其分子机制的广泛应用的工具。然而，可用于单分子荧光测量的标记物种的浓度上限处于低纳米范围，这低于许多酶促反应的米氏常数和许多生物分子的生理浓度。这种差异限制了单分子荧光工具的应用。已经开发了几种技术来突破浓度障碍。在本概念中，我们专注于回顾这些技术的基本原理，并希望激发开发新的更好的工具来实现这一目标。
