Journal Article

Super-resolution imaging of fluorescent dipoles via polarized structured illumination microscopy

Xi, Peng, Xie, Hao, Luo, Sha, Zhanghao, Karl, Dai, Qionghai, Wang, Yiming, Chen, Xingye, Liu, Yiqiong, Shan, Chunyan, Li, Xiangdong, Liu, Wenhui, Jin, Dayong, Gao, Juntao, Zhang, Yan, Li, Meiqi, Wang, Xiao, Chen, Xiaowei

2019

Overview

Fluorescence polarization microscopy images both the intensity and orientation of fluorescent dipoles and plays a vital role in studying molecular structures and dynamics of bio-complexes. However, current techniques remain difficult to resolve the dipole assemblies on subcellular structures and their dynamics in living cells at super-resolution level. Here we report polarized structured illumination microscopy (pSIM), which achieves super-resolution imaging of dipoles by interpreting the dipoles in spatio-angular hyperspace. We demonstrate the application of pSIM on a series of biological filamentous systems, such as cytoskeleton networks and λ-DNA, and report the dynamics of short actin sliding across a myosin-coated surface. Further, pSIM reveals the side-by-side organization of the actin ring structures in the membrane-associated periodic skeleton of hippocampal neurons and images the dipole dynamics of green fluorescent protein-labeled microtubules in live U2OS cells. pSIM applies directly to a large variety of commercial and home-built SIM systems with various imaging modality. Polarization microscopy has been combined with single-molecule localization, but it’s often limited in either speed or resolution. Here the authors present polarized Structured Illumination Microscopy (pSIM), a method that uses polarized laser excitation to measure dye orientation during fast super-resolution live cell imaging.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 14/34 / 14/35 / 14/63 / 631/1647/328/2238 / 639/624/1107/328/1652 / 639/624/1107/328/2238 / Actin / Actins - ultrastructure / Animals / Bacteriophage lambda - genetics / Cell Line, Tumor / Cytoskeleton / Cytoskeleton - ultrastructure / Dipoles / DNA - ultrastructure / Dynamics / Fluorescence / Fluorescence polarization / Green fluorescent protein / Hippocampus / Hippocampus - cytology / Humanities and Social Sciences / Humans / Hyperspaces / Illumination / Image resolution / Kidney / Mice / Microscopy / Microscopy, Fluorescence - methods / Microscopy, Polarization - methods / Microtubules / multidisciplinary / Myosin / Myosins - ultrastructure / Neurons - ultrastructure / Ring structures / Science / Science (multidisciplinary)