Three-dimensional multi-color optical nanoscopy at sub-10-nm resolution based on small-molecule organic probes

Yubing Han; Shijie Tu; Wenwen Gong; Wenli Tao; Mingwei Tang; Yunfei Wei; Cuifang Kuang; Xu Liu; Yu-Hui Zhang; Xiang Hao

doi:10.1016/j.crmeth.2023.100556

Three-dimensional multi-color optical nanoscopy at sub-10-nm resolution based on small-molecule organic probes

Cell Rep Methods. 2023 Sep 25;3(9):100556. doi: 10.1016/j.crmeth.2023.100556. Epub 2023 Aug 14.

Authors

Yubing Han¹, Shijie Tu², Wenwen Gong³, Wenli Tao², Mingwei Tang², Yunfei Wei⁴, Cuifang Kuang⁵, Xu Liu⁶, Yu-Hui Zhang⁷, Xiang Hao⁸

Affiliations

¹ College of Optical Science and Engineering, Zhejiang University, Hangzhou, Zhejiang 310027, China; Britton Chance Center for Biomedical Photonics-MoE Key Laboratory for Biomedical Photonics, Advanced Biomedical Imaging Facility-Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan, Hubei 430074, China. Electronic address: hanyubing@zju.edu.cn.
² College of Optical Science and Engineering, Zhejiang University, Hangzhou, Zhejiang 310027, China.
³ College of Optical Science and Engineering, Zhejiang University, Hangzhou, Zhejiang 310027, China; Institute of Pharmacology, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, Zhejiang 310014, P.R. China.
⁴ Britton Chance Center for Biomedical Photonics-MoE Key Laboratory for Biomedical Photonics, Advanced Biomedical Imaging Facility-Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan, Hubei 430074, China.
⁵ College of Optical Science and Engineering, Zhejiang University, Hangzhou, Zhejiang 310027, China; Ningbo Research Institute, Zhejiang University, Ningbo, Zhejiang 315100, China.
⁶ College of Optical Science and Engineering, Zhejiang University, Hangzhou, Zhejiang 310027, China; Ningbo Research Institute, Zhejiang University, Ningbo, Zhejiang 315100, China. Electronic address: liuxu@zju.edu.cn.
⁷ Britton Chance Center for Biomedical Photonics-MoE Key Laboratory for Biomedical Photonics, Advanced Biomedical Imaging Facility-Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan, Hubei 430074, China. Electronic address: zhangyh@mail.hust.edu.cn.
⁸ College of Optical Science and Engineering, Zhejiang University, Hangzhou, Zhejiang 310027, China. Electronic address: haox@zju.edu.cn.

Abstract

Achieving nanometer-scale resolution remains challenging in expansion microscopy due to photon loss. To address this concern, here we develop a multi-color expansion stimulated emission depletion technique based on small-molecule probes to realize high labeling density and intensity. Our method substantially lowers the barrier to visualizing diverse intracellular proteins and their interactions in three dimensions. It enables us to achieve sub-10-nm resolution in structures such as microfilaments, lysosomes, and mitochondria, providing new insights into cell biology.

Keywords: CP: Imaging, super-resolution imaging, expansion microscopy, small-molecule organic probes, subcellular structures, interactions.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Actin Cytoskeleton
Microscopy*
Mitochondria*