Widengren J, Rigler R
Dept. Medical Biophysics, MBB, Doktorsringen 6A, Karolinska Institute, Stockholm, Sweden.
Cell Mol Biol (Noisy-le-grand). 1998 Jul;44(5):857-79.
Taking advantage of the present day possibilities for ultrasensitive detection by fluorescence, fluorescence correlation spectroscopy (FCS) has over the last ten years emerged as a potentially very powerful technique. In this article we present some results to illustrate the use of FCS for monitoring chemical kinetics on a molecular level and show how, for a wide range of chemical processes, the theoretical treatment can be strongly simplified. The experimental examples given include measurements of ion concentrations and buffer properties, electron transfer reactions, ligand-receptor interactions and diffusion of ligand-receptor complexes in cell membranes. For each of these examples the properties of the FCS technique is discussed in relation to other established techniques used for that particular application. From these examples it is found that FCS can offer important complementary information and, due to the extreme sensitivity of the technique, new information not yet explored by other methods.
利用当今通过荧光进行超灵敏检测的可能性,荧光相关光谱法(FCS)在过去十年中已成为一种潜在的非常强大的技术。在本文中,我们展示了一些结果,以说明FCS在分子水平上监测化学动力学的用途,并展示了对于广泛的化学过程,如何能极大地简化理论处理。给出的实验示例包括离子浓度和缓冲特性的测量、电子转移反应、配体-受体相互作用以及配体-受体复合物在细胞膜中的扩散。对于这些示例中的每一个,都将FCS技术的特性与用于该特定应用的其他既定技术相关联进行了讨论。从这些示例中发现,FCS可以提供重要的补充信息,并且由于该技术的极高灵敏度,还能提供其他方法尚未探索的新信息。