Chen Long, Chen Xingye, Yang Xusan, He Chao, Wang Miaoyan, Xi Peng, Gao Juntao
Department of Automation, Tsinghua University, 100084 Beijing, China.
MOE Key Laboratory of Bioinformatics; Bioinformatics Division, Center for Synthetic & Systems Biology, BNRist; Center for Synthetic & Systems Biology, Tsinghua University, 100084 Beijing, China.
Comput Struct Biotechnol J. 2020 Jun 26;18:2209-2216. doi: 10.1016/j.csbj.2020.06.038. eCollection 2020.
Fluorescence polarization microscopy (FPM) analyzes both intensity and orientation of fluorescence dipole, and reflects the structural specificity of target molecules. It has become an important tool for studying protein organization, orientational order, and structural changes in cells. However, suffering from optical diffraction limit, conventional FPM has low orientation resolution and observation accuracy, as the polarization information is averaged by multiple fluorescent molecules within a diffraction-limited volume. Recently, novel super-resolution FPMs have been developed to break the diffraction barrier. In this review, we will introduce the recent progress to achieve sub-diffraction determination of dipole orientation. Biological applications, based on polarization analysis of fluorescence dipole, are also summarized, with focus on chromophore-target molecule interaction and molecular organization.
荧光偏振显微镜（FPM）可分析荧光偶极子的强度和方向，并反映目标分子的结构特异性。它已成为研究细胞中蛋白质组织、取向顺序和结构变化的重要工具。然而，由于受光学衍射极限的影响，传统的FPM具有较低的取向分辨率和观察精度，因为偏振信息在衍射极限体积内被多个荧光分子平均化了。最近，新型超分辨率FPM已被开发出来以突破衍射屏障。在这篇综述中，我们将介绍在实现偶极子方向的亚衍射测定方面的最新进展。基于荧光偶极子偏振分析的生物学应用也将被总结，重点是发色团-目标分子相互作用和分子组织。
