Li Xuesong

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Last Name
Li
First Name
Xuesong
ORCID
0000-0002-1795-4885

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  • Article
    Three-dimensional structured illumination microscopy with enhanced axial resolution
    (Nature Research, 2023-01-26) Li, Xuesong ; Wu, Yicong ; Su, Yijun ; Rey-Suarez, Ivan ; Matthaeus, Claudia ; Updegrove, Taylor B. ; Wei, Zhuang ; Zhang, Lixia ; Sasaki, Hideki ; Li, Yue ; Guo, Min ; Giannini, John P. ; Vishwasrao, Harshad D. ; Chen, Jiji ; Lee, Shih-Jong J. ; Shao, Lin ; Liu, Huafeng ; Ramamurthi, Kumaran S. ; Taraska, Justin W. ; Upadhyaya, Arpita ; La Riviere, Patrick ; Shroff, Hari
    The axial resolution of three-dimensional structured illumination microscopy (3D SIM) is limited to ∼300 nm. Here we present two distinct, complementary methods to improve axial resolution in 3D SIM with minimal or no modification to the optical system. We show that placing a mirror directly opposite the sample enables four-beam interference with higher spatial frequency content than 3D SIM illumination, offering near-isotropic imaging with ∼120-nm lateral and 160-nm axial resolution. We also developed a deep learning method achieving ∼120-nm isotropic resolution. This method can be combined with denoising to facilitate volumetric imaging spanning dozens of timepoints. We demonstrate the potential of these advances by imaging a variety of cellular samples, delineating the nanoscale distribution of vimentin and microtubule filaments, observing the relative positions of caveolar coat proteins and lysosomal markers and visualizing cytoskeletal dynamics within T cells in the early stages of immune synapse formation.