中文版 | English
Title

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

Author
Corresponding AuthorGao,Juntao
Publication Years
2022-12-01
DOI
Source Title
ISSN
2095-5545
EISSN
2047-7538
Volume11Issue:1
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.
URL[Source Record]
Indexed By
Language
English
SUSTech Authorship
Others
WOS Accession No
WOS:000736928800002
Scopus EID
2-s2.0-85122085156
Data Source
Scopus
Citation statistics
Cited Times [WOS]:0
Document TypeJournal Article
Identifierhttps://kc.sustech.edu.cn/handle/2SGJ60CL/264242
DepartmentCollege of Engineering
工学院_生物医学工程系
Affiliation
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,United States
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,2007,Australia
9.National Biomedical Imaging Center,Peking University,Beijing,100871,China
Recommended Citation
GB/T 7714
Guan,Meiling,Wang,Miaoyan,Zhanghao,Karl,et al. Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells[J]. Light: Science and Applications,2022,11(1).
APA
Guan,Meiling.,Wang,Miaoyan.,Zhanghao,Karl.,Zhang,Xu.,Li,Meiqi.,...&Gao,Juntao.(2022).Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells.Light: Science and Applications,11(1).
MLA
Guan,Meiling,et al."Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells".Light: Science and Applications 11.1(2022).
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