Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells

Meiling Guan; Miaoyan Wang; Karl Zhanghao; Xu Zhang; Meiqi Li; Wenhui Liu; Jing Niu; Xusan Yang; Long Chen; Zhenli Jing; Micheal Q. Zhang; Dayong Jin; Peng Xi; Juntao Gao

doi:10.1038/s41377-021-00689-1

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Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells

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- Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells
- Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells
- LSA 2022年11卷第1期页码：50-62
- Affiliations：
  
  1.Department of Biomedical Engineering, College of Future Technology, Peking University, Beijing 100871, China
  2.UTS-SUStech Joint Research Centre for Biomedical Materials & Devices, Department of Biomedical Engineering, College of Engineering, Southern University of Science and Technology, Shenzhen, Guangdong, China
  3.MOE Key Laboratory of Bioinformatics, Bioinformatics Division, Center for Synthetic & Systems Biology, BNRist, Beijing, China
  4.Center for Synthetic & Systems Biology; Department of Automation, Tsinghua University, Beijing 100084, China
  5.Beijing Institute of Collaborative Innovation, Beijing 100094, China
  6.Department of Biological Sciences and Center for System Biology, The University of Texas at Dallas, Richardson 75080, USA
  7.School of Medical Sciences, Tsinghua University, Beijing 100084, China
  8.Institute for Biomedical Materials and Devices (IBMD), Faculty of Science, University of Technology Sydney, Sydney, NSW 2007, Australia
  9.National Biomedical Imaging Center, Peking University, Beijing 100871, China
- Author bio：
  
  Juntao Gao (jtgao@tsinghua.edu.cn)
- Funds：
- DOI：10.1038/s41377-021-00689-1
  中图分类号：
- 纸质出版日期：2022-01-31，
  
  网络出版日期：2022-01-01，
  
  收稿日期：2021-04-28，
  
  修回日期：2021-11-05，
  
  录用日期：2021-11-27
- Accepted：
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Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells[J]. LSA, 2022,11(1):50-62.

Guan, M. et al. Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells. Light: Science & Applications, 11, 50-62 (2022).
Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells[J]. LSA, 2022,11(1):50-62. DOI： 10.1038/s41377-021-00689-1.

Guan, M. et al. Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells. Light: Science & Applications, 11, 50-62 (2022). DOI： 10.1038/s41377-021-00689-1.

摘要

Abstract

The orientation of fluorophores can reveal crucial information about the structure and dynamics of their associated subcellular organelles. Despite significant progress in super-resolution

fluorescence polarization microscopy remains limited to unique samples with relatively strong polarization modulation and not applicable to the weak polarization signals in samples due to the excessive background noise. Here we apply optical lock-in detection to amplify the weak polarization modulation with super-resolution. This novel technique

termed optical lock-in detection super-resolution dipole orientation mapping (OLID-SDOM)

could achieve a maximum of 100 frames per second and rapid extraction of 2D orientation

and distinguish distance up to 50 nm

making it suitable for monitoring structural dynamics concerning orientation changes in vivo. OLID-SDOM was employed to explore the universal anisotropy of a large variety of GFP-tagged subcellular organelles

including mitochondria

lysosome

Golgi

endosome

etc. We found that OUF (Orientation Uniformity Factor) of OLID-SDOM can be specific for different subcellular organelles

indicating that the anisotropy was related to the function of the organelles

and OUF can potentially be an indicator to distinguish normal and abnormal cells (even cancer cells). Furthermore

dual-color super-resolution OLID-SDOM imaging of lysosomes and actins demonstrates its potential in studying dynamic molecular interactions. The subtle anisotropy changes of expanding and shrinking dendritic spines in live neurons were observed with real-time OLID-SDOM. Revealing previously unobservable fluorescence anisotropy in various samples and indicating their underlying dynamic molecular structural changes

OLID-SDOM expands the toolkit for live cell research.

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