(Peer-Reviewed) Multiplexed stimulated emission depletion nanoscopy (mSTED) for 5-color live-cell long-term imaging of organelle interactome
Yuran Huang 黄宇然 ¹, Zhimin Zhang 张智敏 ¹ ², Wenli Tao 陶汶理 ¹, Yunfei Wei 魏云飞 ³, Liang Xu 徐良 ¹, Wenwen Gong 宫文文 ¹, Jiaqiang Zhou 周嘉强 ⁴, Liangcai Cao 曹良才 ⁵, Yong Liu 刘勇 ⁶, Yubing Han 韩于冰 ¹ ³, Cuifang Kuang 匡翠方 ¹ ² ⁷, Xu Liu 刘旭 ¹ ²
¹ State Key Laboratory of Extreme Photonics and Instrumentation, College of Optical Science and Engineering, Zhejiang University, Hangzhou 310027, China
中国 杭州 浙江大学光电科学与工程学院 极端光学技术与仪器全国重点实验室
² Research Center for Intelligent Chips and Devices, Zhejiang Lab, Hangzhou 311121, China
中国 杭州 之江实验室 智能芯片与器件研究中心
³ Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, China
中国 武汉 华中科技大学武汉光电国家研究中心
⁴ Department of Endocrinology and Metabolism, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310016, China
中国 杭州 浙江大学医学院附属邵逸夫医院 内分泌代谢科
⁵ Department of Precision Instruments, Tsinghua University, Beijing 100084, China
中国 北京 清华大学精密仪器系
⁶ College of electronics and information engineering, Shanghai University of Electrical Power, Shanghai 200090, China
中国 上海 上海电力大学 电子与信息工程学院
⁷ ZJU-Hangzhou Global Scientific and Technological Innovation Center, Hangzhou 311200, China
中国 杭州 浙江大学杭州国际科创中心
Opto-Electronic Advances, 2024-07-05
Abstract
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.
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Hanyu Jin, Hao Li, Zhuolin Chen, Liangye Li, Zurui Wang, Kai Zhou, Weijian Hang, Zhipeng Chen, Junfeng Chen, Kai Shen, Jianfeng Wen, Chen Chen, Cunzheng Fan, Zhijun Yan, Feng Wang, Qizhen Sun
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Opto-Electronic Advances
2026-07-10
