Multiplexed stimulated emission depletion nanoscopy (mSTED) for 5-color live-cell long-term imaging of organelle interactome

Yuran Huang; Zhimin Zhang; Wenli Tao; Yunfei Wei; Liang Xu; Wenwen Gong; Jiaqiang Zhou; Liangcai Cao; Yong Liu; Yubing Han; Cuifang Kuang; Xu Liu

doi:10.29026/oea.2024.240035

Opto-Electronic Advances (Jul 2024)

Multiplexed stimulated emission depletion nanoscopy (mSTED) for 5-color live-cell long-term imaging of organelle interactome

Yuran Huang,
Zhimin Zhang,
Wenli Tao,
Yunfei Wei,
Liang Xu,
Wenwen Gong,
Jiaqiang Zhou,
Liangcai Cao,
Yong Liu,
Yubing Han,
Cuifang Kuang,
Xu Liu

Affiliations

Yuran Huang: State Key Laboratory of Extreme Photonics and Instrumentation, College of Optical Science and Engineering, Zhejiang University, Hangzhou 310027, China
Zhimin Zhang: State Key Laboratory of Extreme Photonics and Instrumentation, College of Optical Science and Engineering, Zhejiang University, Hangzhou 310027, China
Wenli Tao: State Key Laboratory of Extreme Photonics and Instrumentation, College of Optical Science and Engineering, Zhejiang University, Hangzhou 310027, China
Yunfei Wei: Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, China
Liang Xu: State Key Laboratory of Extreme Photonics and Instrumentation, College of Optical Science and Engineering, Zhejiang University, Hangzhou 310027, China
Wenwen Gong: State Key Laboratory of Extreme Photonics and Instrumentation, College of Optical Science and Engineering, Zhejiang University, Hangzhou 310027, China
Jiaqiang Zhou: Department of Endocrinology and Metabolism, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310016, China
Liangcai Cao: Department of Precision Instruments, Tsinghua University, Beijing 100084, China
Yong Liu: College of electronics and information engineering, Shanghai University of Electrical Power, Shanghai 200090, China
Yubing Han: State Key Laboratory of Extreme Photonics and Instrumentation, College of Optical Science and Engineering, Zhejiang University, Hangzhou 310027, China
Cuifang Kuang: State Key Laboratory of Extreme Photonics and Instrumentation, College of Optical Science and Engineering, Zhejiang University, Hangzhou 310027, China
Xu Liu: State Key Laboratory of Extreme Photonics and Instrumentation, College of Optical Science and Engineering, Zhejiang University, Hangzhou 310027, China

DOI: https://doi.org/10.29026/oea.2024.240035
Journal volume & issue: Vol. 7, no. 7
pp. 1 – 10

Abstract

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Stimulated emission depletion microscopy (STED) holds great potential in biological science applications, especially in studying nanoscale subcellular structures. However, multi-color STED imaging in live-cell remains challenging due to the limited excitation wavelengths and large amount of laser radiation. Here, we develop a multiplexed live-cell STED method to observe more structures simultaneously with limited photo-bleaching and photo-cytotoxicity. By separating live-cell fluorescent probes with similar spectral properties using phasor analysis, our method enables five-color live-cell STED imaging and reveals long-term interactions between different subcellular structures. The results here provide an avenue for understanding the complex and delicate interactome of subcellular structures in live-cell.

Published in Opto-Electronic Advances

ISSN: 2096-4579 (Print)
Publisher: Institue of Optics and Electronics, Chinese Academy of Sciences
Country of publisher: China
LCC subjects: Science: Physics: Optics. Light
Website: http://www.oejournal.org/oea

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