Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells
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 polariz...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Publishing Group
2022-01-01
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| Series: | Light: Science & Applications |
| Online Access: | https://doi.org/10.1038/s41377-021-00689-1 |
| _version_ | 1819275566864400384 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Meiling Guan |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-23T23:26:22Z |
| format | Article |
| id | doaj.art-1255444c548941db9fc467c4c7778706 |
| institution | Directory Open Access Journal |
| issn | 2047-7538 |
| language | English |
| last_indexed | 2024-12-23T23:26:22Z |
| publishDate | 2022-01-01 |
| publisher | Nature Publishing Group |
| record_format | Article |
| series | Light: Science & Applications |
| spelling | doaj.art-1255444c548941db9fc467c4c77787062022-12-21T17:26:13ZengNature Publishing GroupLight: Science & Applications2047-75382022-01-0111111310.1038/s41377-021-00689-1Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cellsMeiling Guan0Miaoyan Wang1Karl Zhanghao2Xu Zhang3Meiqi Li4Wenhui Liu5Jing Niu6Xusan Yang7Long Chen8Zhenli Jing9Micheal Q. Zhang10Dayong Jin11Peng Xi12Juntao Gao13Department of Biomedical Engineering, College of Future Technology, Peking UniversityDepartment of Biomedical Engineering, College of Future Technology, Peking UniversityDepartment of Biomedical Engineering, College of Future Technology, Peking UniversityMOE Key Laboratory of Bioinformatics, Bioinformatics Division, Center for Synthetic & Systems Biology, BNRistDepartment of Biomedical Engineering, College of Future Technology, Peking UniversityCenter for Synthetic & Systems Biology; Department of Automation, Tsinghua UniversityMOE Key Laboratory of Bioinformatics, Bioinformatics Division, Center for Synthetic & Systems Biology, BNRistDepartment of Biomedical Engineering, College of Future Technology, Peking UniversityMOE Key Laboratory of Bioinformatics, Bioinformatics Division, Center for Synthetic & Systems Biology, BNRistMOE Key Laboratory of Bioinformatics, Bioinformatics Division, Center for Synthetic & Systems Biology, BNRistMOE Key Laboratory of Bioinformatics, Bioinformatics Division, Center for Synthetic & Systems Biology, BNRistUTS-SUStech Joint Research Centre for Biomedical Materials & Devices, Department of Biomedical Engineering, College of Engineering, Southern University of Science and TechnologyDepartment of Biomedical Engineering, College of Future Technology, Peking UniversityMOE Key Laboratory of Bioinformatics, Bioinformatics Division, Center for Synthetic & Systems Biology, BNRistAbstract 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.https://doi.org/10.1038/s41377-021-00689-1 |
| spellingShingle | 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 Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells Light: Science & Applications |
| title | Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells |
| title_full | Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells |
| title_fullStr | Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells |
| title_full_unstemmed | Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells |
| title_short | Polarization modulation with optical lock-in detection reveals universal fluorescence anisotropy of subcellular structures in live cells |
| title_sort | polarization modulation with optical lock in detection reveals universal fluorescence anisotropy of subcellular structures in live cells |
| url | https://doi.org/10.1038/s41377-021-00689-1 |
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