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

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外文摘要：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.

外文关键词：

optical nanoscopyphasor analysismulticolor live cell imaging

作者：

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

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作者单位：

State Key Laboratory of Extreme Photonics and Instrumentation,College of Optical Science and Engineering,Zhejiang University,Hangzhou 310027,China

Research Center for Intelligent Chips and Devices,Zhejiang Lab,Hangzhou 311121,China

Wuhan National Laboratory for Optoelectronics,Huazhong University of Science and Technology,Wuhan 430074,China

Department of Endocrinology and Metabolism,Sir Run Run Shaw Hospital,Zhejiang University School of Medicine,Hangzhou 310016,China

Department of Precision Instruments,Tsinghua University,Beijing 100084,China

College of electronics and information engineering,Shanghai University of Electrical Power,Shanghai 200090,China

ZJU-Hangzhou Global Scientific and Technological Innovation Center,Hangzhou 311200,China

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基金：

following grants:National Natural Science Foundation of Chinafollowing grants:National Natural Science Foundation of ChinaSTI 2030-Major ProjectsFundamental Research Funds for the Central UniversitiesOpen Project Program of Wuhan National Laboratory for Optoelectronics

项目编号：

62125504623611666312021ZD0200401226-2022-002012021WNLOKF007

出版年：

2024

DOI：

10.29026/oea.2024.240035

光电进展(英文版)

ISSN：

年,卷(期)：2024.7(7)