Accurate 3D single-molecule localization via vectorial in situ point spread function retrieval and aberration assessment  

作　　者：XINXUN YANG HONGFEI ZHU YILE SUN HANMENG WU YUBING HAN XIANG HAO 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]Department of Biomedical Engineering,The Chinese University of Hong Kong,Hong Kong,China [3]ZJU-Hangzhou Global Scientific and Technological Innovation Center,Hangzhou 311200,China [4]Collaborative Innovation Center of Extreme Optics,Shanxi University,Taiyuan 030006,China

出　　处：《Photonics Research》2024年第11期2447-2461,共15页光子学研究(英文版)

基　　金：National Natural Science Foundation of China(62125504,62361166631);STI 2030—Major Projects(2021ZD0200401);Fundamental Research Funds for the Central Universities(2022FZZX01-20);Open Project Program of Wuhan National Laboratory for Optoelectronics(2021WNLOKF007)。

摘　　要：Single-molecule localization microscopy(SMLM)gradually plays an important role in deep tissue imaging.However,current SMLM methods primarily rely on fiducial marks,neglecting aberrations introduced by thick samples,thereby resulting in decreased image quality in deep tissues.Here,we introduce vectorial in situ point spread function(PSF)retrieval(VISPR),a method that retrieves a precise PSF model considering both systemand sample-induced aberrations under SMLM conditions.By employing the theory of vectorial PSF model and maximum likelihood estimation(MLE)phase retrieval,VISPR is capable of reconstructing an accurate in situ 3D PSF model achieving the theoretically minimum uncertainty and accurately reflecting three-dimensional information of single molecules.This capability enables accurate 3D super-resolution reconstruction in deep regions away from the coverslips.Additionally,VISPR demonstrates applicability in low signal-to-noise ratio scenarios and compatibility with various SMLM microscope modalities.From both simulations and experiments,we verified the superiority and effectiveness of VISPR.We anticipate that VISPR will become a pivotal tool for advancing deep tissue SMLM imaging.

关 键 词：function thereby PRECISE 

分 类 号：O35[理学—流体力学]