PhotoniX, Volume. 5, Issue 1, 12(2024)

Three-dimensional dipole orientation mapping with high temporal-spatial resolution using polarization modulation

Suyi Zhong1...2, Liang Qiao3, Xichuan Ge3, Xinzhu Xu1,2, Yunzhe Fu1,2, Shu Gao4, Karl Zhanghao1, Huiwen Hao5, Wenyi Wang1,2, Meiqi Li6,*, and Peng Xi12,** |Show fewer author(s)
Author Affiliations
  • 1Department of Biomedical Engineering, College of Future Technology, Peking University, Beijing 100871, China
  • 2National Biomedical Imaging Center, Peking University, Beijing 100871, China
  • 3Airy Technologies Co., Ltd., Beijing, China
  • 4Institute of Molecular Medicine, National Biomedical Imaging Center, College of Future Technology, Peking University, Beijing 100871, China
  • 5Standard Imaging Company, Beijing, China
  • 6School of Life Sciences, Peking University, Beijing 100871, China
  • show less

    Fluorescence polarization microscopy is widely used in biology for molecular orientation properties. However, due to the limited temporal resolution of single-molecule orientation localization microscopy and the limited orientation dimension of polarization modulation techniques, achieving simultaneous high temporal-spatial resolution mapping of the three-dimensional (3D) orientation of fluorescent dipoles remains an outstanding problem. Here, we present a super-resolution 3D orientation mapping (3DOM) microscope that resolves 3D orientation by extracting phase information of the six polarization modulation components in reciprocal space. 3DOM achieves an azimuthal precision of 2° and a polar precision of 3° with spatial resolution of up to 128 nm in the experiments. We validate that 3DOM not only reveals the heterogeneity of the milk fat globule membrane, but also elucidates the 3D structure of biological filaments, including the 3D spatial conformation of λ-DNA and the structural disorder of actin filaments. Furthermore, 3DOM images the dipole dynamics of microtubules labeled with green fluorescent protein in live U2OS cells, reporting dynamic 3D orientation variations. Given its easy integration into existing wide-field microscopes, we expect the 3DOM microscope to provide a multi-view versatile strategy for investigating molecular structure and dynamics in biological macromolecules across multiple spatial and temporal scales.

    Tools

    Get Citation

    Copy Citation Text

    Suyi Zhong, Liang Qiao, Xichuan Ge, Xinzhu Xu, Yunzhe Fu, Shu Gao, Karl Zhanghao, Huiwen Hao, Wenyi Wang, Meiqi Li, Peng Xi. Three-dimensional dipole orientation mapping with high temporal-spatial resolution using polarization modulation[J]. PhotoniX, 2024, 5(1): 12

    Download Citation

    EndNote(RIS)BibTexPlain Text
    Save article for my favorites
    Paper Information

    Category: Research Articles

    Received: Dec. 13, 2023

    Accepted: Mar. 28, 2024

    Published Online: Jan. 23, 2025

    The Author Email: Li Meiqi (limeiqi@pku.edu.cn), Xi Peng (xipeng@pku.edu.cn)

    DOI:10.1186/s43074-024-00127-6

    Topics