Beijing National Laboratory for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China and The University of Chinese Academy of Sciences, Beijing 100049, China.
J Chem Phys. 2020 Nov 7;153(17):170903. doi: 10.1063/5.0024324.
It is well documented that conventional methods such as dynamic light scattering have encountered difficulties in characterizing charged macromolecules and, therefore, it is desirable that new methods and techniques are introduced. With the ultra-high sensitivity, single molecule fluorescence spectroscopy has successfully lowered the detection limit considerably and enabled measurement under extreme dilution conditions-around the concentration of 10M-at which the effect of inter-chain electrostatic repulsion is suppressed. Furthermore, the excellent spatial and temporal resolution as well as the capacity of molecular recognition of these methods help in obtaining rich information of charged macromolecules. This paper summarizes the applications of single molecule fluorescence spectroscopy, especially fluorescence correlation spectroscopy and photon counting histogram, in the studies on charged macromolecules in aqueous solutions and plenty of new information has been revealed on the molecular conformation, counterion distribution, and a few important governing factors. The powerfulness and effectiveness of single molecule fluorescence spectroscopy make it promising in the investigations of charged macromolecules.
有大量文献记载表明,传统方法(如动态光散射)在表征带电大分子时遇到了困难,因此,有必要引入新的方法和技术。单分子荧光光谱具有超高灵敏度,成功地大大降低了检测限,并能够在极端稀释条件下进行测量——在这种浓度下,链间静电排斥的影响被抑制。此外,这些方法具有出色的空间和时间分辨率以及分子识别能力,有助于获取有关带电大分子的丰富信息。本文总结了单分子荧光光谱学,特别是荧光相关光谱学和光子计数直方图在水溶液中带电大分子研究中的应用,并揭示了有关分子构象、抗衡离子分布以及一些重要控制因素的大量新信息。单分子荧光光谱学的强大功能使其有望用于研究带电大分子。