In Nature methods ; h5-index 152.0
Single-molecule localization microscopy in a typical wide-field setup has been widely used for investigating subcellular structures with super resolution; however, field-dependent aberrations restrict the field of view (FOV) to only tens of micrometers. Here, we present a deep-learning method for precise localization of spatially variant point emitters (FD-DeepLoc) over a large FOV covering the full chip of a modern sCMOS camera. Using a graphic processing unit-based vectorial point spread function (PSF) fitter, we can fast and accurately model the spatially variant PSF of a high numerical aperture objective in the entire FOV. Combined with deformable mirror-based optimal PSF engineering, we demonstrate high-accuracy three-dimensional single-molecule localization microscopy over a volume of ~180 × 180 × 5 μm3, allowing us to image mitochondria and nuclear pore complexes in entire cells in a single imaging cycle without hardware scanning; a 100-fold increase in throughput compared to the state of the art.
Fu Shuang, Shi Wei, Luo Tingdan, He Yingchuan, Zhou Lulu, Yang Jie, Yang Zhichao, Liu Jiadong, Liu Xiaotian, Guo Zhiyong, Yang Chengyu, Liu Chao, Huang Zhen-Li, Ries Jonas, Zhang Mingjie, Xi Peng, Jin Dayong, Li Yiming
2023-Feb-23
