Scanning Switch-off Microscopy for Super-Resolution Fluorescence Imaging

作者：Zhaoshuai Gao, Shangguo Hou, Suhui Deng, Le Liang, Fei Wang, Linjie Guo, Weina Fang, Qian Li, Bin Kang, Hong‐Yuan Chen, Chunhai Fan · 发表于：Nano Letters · 年份：2024 · DOI：10.1021/acs.nanolett.4c02452 · 被引用次数：7 · 研究领域：Advanced Fluorescence Microscopy Techniques、Cell Image Analysis Techniques、Advanced Electron Microscopy Techniques and Applications

Super-resolution (SR) microscopy provides a revolutionary optical imaging approach by breaking the diffraction limit of light, while the commonly required special instrumentation with complex optical setup hampers its popularity. Here, we present a scanning switch-off microscopy (SSM) concept that exploits the omnipresent switch-off response of fluorophores to enable super-resolution imaging using a commercial confocal microscope. We validated the SSM model with theoretical calculations and experiments. An imaging resolution of ∼100 nm was obtained for DNA origami nanostructures and cellular cytoskeletons using fluorescent labels of Alexa 405, Alexa 488, Cy3, and Atto 488. Notably, super-resolution imaging of live cells was realized with SSM, by employing a dronpa fluorescent protein as the fluorescent label. In principle, this SSM method can be applied to any excitation laser scanning-based microscope.