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

*Corresponding author for this work

Research output: Contribution to journalArticle

55 Citations (Scopus)
78 Downloads (Pure)

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.

Original languageEnglish
Article numbere16166
Pages (from-to)1-8
Number of pages8
JournalLight: Science and Applications
Volume5
Issue number10
DOIs
Publication statusPublished - 1 Oct 2016
Externally publishedYes

Bibliographical note

Copyright the Author(s) 2016. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

Keywords

  • dipole
  • fluorescence polarization microscopy
  • orientation mapping
  • polarization modulation
  • super-resolution

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  • Cite this

    Zhanghao, K., Chen, L., Yang, X. S., Wang, M. Y., Jing, Z. L., Han, H. B., ... Xi, P. (2016). Super-resolution dipole orientation mapping via polarization demodulation. Light: Science and Applications, 5(10), 1-8. [e16166]. https://doi.org/10.1038/lsa.2016.166