Ultra-high spatio-temporal resolution imaging with parallel acquisition-readout structured illumination microscopy(PAR-SIM)  

作　　者：Xinzhu Xu Wenyi Wang Liang Qiao Yunzhe Fu Xichuan Ge Kun Zhao Karl Zhanghao Meiling Guan Xin Chen Meiqi Li Dayong Jin Peng Xi 

机构地区：[1]Department of Biomedical Engineering,College of Future Technology,Peking University,Beijing,100871,China [2]Wallace H.Coulter Dept.of Biomedical Engineering,Georgia Institute of Technology and Emory University,Atlanta,30332,GA,USA [3]National Biomedical Imaging Center,Peking University,Beijing,100871,China [4]Department of Biomedical Engineering,College of Engineering,Southern University of Science and Technology,Shenzhen,518055,Guangdong,China [5]Airy Technologies Co.,Ltd.,Beijing,100086,China [6]Eastern Institute for Advanced Study,Eastern Institute of Technology,Ningbo,Zhejiang,315200,China [7]School of Life Science,Peking University,Beijing,100871,China [8]Institute for Biomedical Materials and Devices(IBMD),Faculty of Science,University of Technology Sydney,Sydney,NSW,2007,Australia

出　　处：《Light(Science & Applications)》2024年第6期1140-1158,共19页光（科学与应用)（英文版）

基　　金：supported by the National Key R&D Program of China (2022YFC3401100);the National Natural Science Foundation of China (62025501,92150301,62335008).

摘　　要：Structured illumination microscopy(SIM)has emerged as a promising super-resolution fluorescence imaging technique,offering diverse configurations and computational strategies to mitigate phototoxicity during real-time imaging of biological specimens.Traditional efforts to enhance system frame rates have concentrated on processing algorithms,like rolling reconstruction or reduced frame reconstruction,or on investments in costly sCMOS cameras with accelerated row readout rates.In this article,we introduce an approach to elevate SIM frame rates and region of interest(ROI)coverage at the hardware level,without necessitating an upsurge in camera expenses or intricate algorithms.Here,parallel acquisition-readout SIM(PAR-SIM)achieves the highest imaging speed for fluorescence imaging at currently available detector sensitivity.By using the full frame-width of the detector through synchronizing the pattern generation and image exposure-readout process,we have achieved a fundamentally stupendous information spatial-temporal flux of 132.9 MPixels·s^(−1),9.6-fold that of the latest techniques,with the lowest SNR of−2.11 dB and 100 nm resolution.PAR-SIM demonstrates its proficiency in successfully reconstructing diverse cellular organelles in dual excitations,even under conditions of low signal due to ultra-short exposure times.Notably,mitochondrial dynamic tubulation and ongoing membrane fusion processes have been captured in live COS-7 cell,recorded with PAR-SIM at an impressive 408 Hz.We posit that this novel parallel exposure-readout mode not only augments SIM pattern modulation for superior frame rates but also holds the potential to benefit other complex imaging systems with a strategic controlling approach.

关 键 词：hardware ILLUMINATION RESOLUTION 

分 类 号：TH742[机械工程—光学工程]