Super-resolution dipole orientation mapping via polarization demodulation  

作　　者：Karl Zhanghao Long Chen Xu-San Yang Miao-Yan Wang Zhen-Li Jing Hong-Bin Han Michael Q Zhang Dayong Jin Jun-Tao Gao Peng Xi 

机构地区：[1]Department of Biomedical Engineering,College of Engineering,Peking University,Beijing 100871,China [2]Department of Automation,Tsinghua University,Beijing 100084,China [3]Bioinformatics Division,TNLIST,MOE Key Laboratory of Bioinformatics and Center for Synthetic&Systems Biology,Tsinghua University,Beijing 100084,China [4]Department of Radiology,Peking University Third Hospital,Beijing 100191,China [5]Department of Biological Sciences,Center for Systems Biology,The University of Texas,Dallas 800 West Campbell Road,RL11,Richardson,TX 75080-3021,USA [6]Department of Basic Medical Sciences,School of Medicine,Tsinghua University,Beijing 100084,China [7]Institute for Biomedical Materials and Devices(IBMD),Faculty of Science,University of Technology Sydney,NSW 2007,Australia

出　　处：《Light(Science & Applications)》2016年第1期112-119,共8页光（科学与应用)（英文版）

基　　金：supported by the National Key Basic Research Program(973 Program,2012CB316503);the National Instrument Development Special Program(2013YQ03065102);the National Natural Science Foundation of China(31361163004,31327901,61475010 and 61428501);supported by UTD funds.

摘　　要：Fluorescence polarization microscopy(FPM)aims to detect the dipole orientation of fluorophores and to resolve structural information for labeled organelles via wide-field or confocal microscopy.Conventional FPM often suffers from the presence of a large number of molecules within the diffraction-limited volume,with averaged fluorescence polarization collected from a group of dipoles with different orientations.Here,we apply sparse deconvolution and least-squares estimation to fluorescence polarization modulation data and demonstrate a super-resolution dipole orientation mapping(SDOM)method that resolves the effective dipole orientation from a much smaller number of fluorescent molecules within a sub-diffraction focal area.We further apply this method to resolve structural details in both fixed and live cells.For the first time,we show that different borders of a dendritic spine neck exhibit a heterogeneous distribution of dipole orientation.Furthermore,we illustrate that the dipole is always perpendicular to the direction of actin filaments in mammalian kidney cells and radially distributed in the hourglass structure of the septin protein under specific labelling.The accuracy of the dipole orientation can be further mapped using the orientation uniform factor,which shows the superiority of SDOM compared with its wide-field counterpart as the number of molecules is decreased within the smaller focal area.Using the inherent feature of the orientation dipole,the SDOM technique,with its fast imaging speed(at sub-second scale),can be applied to a broad range of fluorescently labeled biological systems to simultaneously resolve the valuable dipole orientation information with super-resolution imaging.

关 键 词：DIPOLE fluorescence polarization microscopy orientation mapping polarization modulation SUPER-RESOLUTION 

分 类 号：TG1[金属学及工艺—金属学]