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

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 

机构地区：[1]State Key Laboratory of Extreme Photonics and Instrumentation,College of Optical Science and Engineering,Zhejiang University,Hangzhou 310027,China [2]Research Center for Intelligent Chips and Devices,Zhejiang Lab,Hangzhou 311121,China [3]Wuhan National Laboratory for Optoelectronics,Huazhong University of Science and Technology,Wuhan 430074,China [4]Department of Endocrinology and Metabolism,Sir Run Run Shaw Hospital,Zhejiang University School of Medicine,Hangzhou 310016,China [5]Department of Precision Instruments,Tsinghua University,Beijing 100084,China [6]College of electronics and information engineering,Shanghai University of Electrical Power,Shanghai 200090,China [7]ZJU-Hangzhou Global Scientific and Technological Innovation Center,Hangzhou 311200,China

出　　处：《Opto-Electronic Advances》2024年第7期17-26,共10页光电进展（英文）

基　　金：supported by the following grants:National Natural Science Foundation of China(62125504,62361166631);STI 2030-Major Projects(2021ZD0200401);the Fundamental Research Funds for the Central Universities(226-2022-00201);the Open Project Program of Wuhan National Laboratory for Optoelectronics(2021WNLOKF007).

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.

关 键 词：optical nanoscopy phasor analysis multicolor live cell imaging 

分 类 号：Q2-33[生物学—细胞生物学] TP391.41[自动化与计算机技术—计算机应用技术]