achieving λ10 resolution cw sted nanoscopy with a tisapphire oscillator实现λ10决议cw、nanoscopy tisapphire振荡器.pdfVIP

Achieving l/10 Resolution CW STED Nanoscopy with a Ti:Sapphire Oscillator 1,2 2 3 5 3 4 4 Yujia Liu , Yichen Ding , Eric Alonas , Wenli Zhao , Philip J. Santangelo , Dayong Jin , James A. Piper , 5 1,2 2 Junlin Teng , Qiushi Ren , Peng Xi * 1 School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, China, 2 Department of Biomedical Engineering, College of Engineering, Peking University, Beijing, China, 3 Wallace H Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia, United States of America, 4 Advanced Cytometry Labs, MQphotonics Research Centre, Macquarie University, Sydney, New South Wales, Australia, 5 College of Life Sciences, Peking University, Beijing, China Abstract In this report, a Ti:Sapphire oscillator was utilized to realize synchronization-free stimulated emission depletion (STED) microscopy. With pump power of 4.6 W and sample irradiance of 310 mW, we achieved super-resolution as high as 71 nm. With synchronization-free STED, we imaged 200 nm nanospheres as well as all three cytoskeletal elements (microtubules, intermediate filaments, and actin filaments), clearly demonstrating the resolving power of synchronization-free STED over conventional diffraction limited imaging. It also allowed us to discover that, Dylight 650, exhibits improved performance over ATTO647N, a fluorophore frequently used in STED. Furthermore, we applied synchronization-free STED to image fluorescently-labeled intracellular viral RNA granules, which otherwise cannot be differentiated by confocal microscopy. Thanks to the wi