Elucidating subcellular architecture and dynamics at isotropic 100-nm resolution with 4Pi-SIM
Three-dimensional structured illumination microscopy (3D-SIM) provides excellent optical sectioning and doubles the resolution in all dimensions compared with wide-field microscopy. However, its much lower axial resolution results in blurred fine details in that direction and overall image distortion. Here we present 4Pi-SIM, a substantial revamp of I<sup>5</sup>S that synergizes 3D-SIM with interferometric microscopy to achieve isotropic optical resolution through interference in both the illumination and detection wavefronts. We evaluate the performance of 4Pi-SIM by imaging various subcellular structures across different cell types with high fidelity. Furthermore, we demonstrate its capability by conducting time-lapse volumetric imaging over hundreds of time points, achieving a 3D resolution of approximately 100 nm. Additionally, we illustrate its ability to simultaneously image in two colors and capture the rapid interactions between closely positioned organelles in three dimensions. These results underscore the great potential of 4Pi-SIM for elucidating subcellular architecture and revealing dynamic behaviors at the nanoscale.
Authors
- Wen Gou (ORCID: https://orcid.org/0000-0002-1795-2978)
- Yongdeng Zhang (ORCID: https://orcid.org/0000-0002-4569-8137)
- Yanqin Chen (ORCID: https://orcid.org/0000-0002-3302-7643)
- Changliang Liu (ORCID: https://orcid.org/0009-0005-8201-0871)
- Lin Shao (ORCID: https://orcid.org/0000-0002-1905-4442)
- Liangyi Chen (ORCID: https://orcid.org/0000-0003-1270-7321)
- Junchao Fan (ORCID: https://orcid.org/0000-0003-2586-7973)
- Zijing Yu (ORCID: https://orcid.org/0009-0005-1697-3872)
- Bin Xiao (ORCID: https://orcid.org/0000-0001-8469-5302)
- Xiuli Bi (ORCID: https://orcid.org/0000-0003-3134-217X)
- Xiaoyu Li (ORCID: https://orcid.org/0009-0006-9840-6704)
- Qiuzhong Dai (ORCID: https://orcid.org/0000-0001-5490-9816)
- Xiaoshuai Huang (ORCID: https://orcid.org/0000-0002-0495-4720)
- Qian Wang (ORCID: https://orcid.org/0000-0002-0326-4410)
- Xiaochun Yu
- Muyuan Tang
- Bei Zheng (ORCID: https://orcid.org/0000-0003-0575-1607)
- Linlin Chen (ORCID: https://orcid.org/0000-0003-3977-5279)
- Zheyuan Ouyang (ORCID: https://orcid.org/0009-0009-3176-1471)
- Chenqi Ping
Institutions
- Chongqing University of Posts and Telecommunications (CN)
- McGovern Institute for Brain Research (US)
- Peking University (CN)
- Westlake University (CN)
- Yale University (US)
- Peking University Cancer Hospital (CN)
- Chinese Institute for Brain Research (CN)
- Beijing National Laboratory for Molecular Sciences (CN)
- Ministry of Education (ET)
- Center for Life Sciences (CN)
Publication Details
- Journal
- Nature Methods
- Published
- 2024-12-23
- DOI
- https://doi.org/10.1038/s41592-024-02515-z
- Citations
- 22
- Primary Topic
- Advanced Fluorescence Microscopy Techniques
- Type
- article
- Field-Weighted Citation Impact
- 14.57
Funders
- National Science Foundation
- American Occupational Therapy Foundation
- Yale University
- Westlake University
- National Natural Science Foundation of China
- Chinese Academy of Sciences
- Peking University
- Beijing Nova Program
- National Key Research and Development Program of China
- Institute of Biophysics, Chinese Academy of Sciences