Super-resolution dipole orientation mapping via polarization demodulation

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

doi:10.1038/lsa.2016.166

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Super-resolution dipole orientation mapping via polarization demodulation

Original Articles

- Super-resolution dipole orientation mapping via polarization demodulation
- Super-resolution dipole orientation mapping via polarization demodulation
- LSA 2016年5卷第4期
- Affiliations：
  
  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
- Author bio：
  
  D Jin, Email: dayong.jin@uts.edu.au
  JT Gao, Email: jtgao@biomed.tsinghua.edu.cn
  P Xi, Email: xipeng@pku.edu.cn
- Funds：
- DOI：10.1038/lsa.2016.166
  中图分类号：
- 纸质出版日期：2016-12，
  
  网络出版日期：2016-10-21，
  
  收稿日期：2015-12-26，
  
  修回日期：2016-05-09，
  
  录用日期：2016-05-17
- Accepted：
Scan QR Code
Super-resolution dipole orientation mapping via polarization demodulation[J]. LSA, 2016,5(4).

Zhanghao, K. et al. Super-resolution dipole orientation mapping via polarization demodulation. Light: Science & Applications, 5, e16166 (2016).
Super-resolution dipole orientation mapping via polarization demodulation[J]. LSA, 2016,5(4). DOI： 10.1038/lsa.2016.166.

Zhanghao, K. et al. Super-resolution dipole orientation mapping via polarization demodulation. Light: Science & Applications, 5, e16166 (2016). DOI： 10.1038/lsa.2016.166.

摘要

Abstract

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

Keywords

dipolefluorescence polarization microscopyorientation mappingpolarization modulationsuper-resolution

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