Ultra-high spatio-temporal resolution imaging with parallel acquisition-readout structured illumination microscopy(PAR-SIM)OACSTPCD
Ultra-high spatio-temporal resolution imaging with parallel acquisition-readout structured illumination microscopy(PAR-SIM)
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 100nm 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.
Xinzhu Xu;Meiqi Li;Dayong Jin;Peng Xi;Wenyi Wang;Liang Qiao;Yunzhe Fu;Xichuan Ge;Kun Zhao;Karl Zhanghao;Meiling Guan;Xin Chen
Department of Biomedical Engineering,College of Future Technology,Peking University,Beijing 100871,China||Wallace H.Coulter Dept.of Biomedical Engineering,Georgia Institute of Technology and Emory University,Atlanta 30332 GA,USA||National Biomedical Imaging Center,Peking University,Beijing 100871,China||Department of Biomedical Engineering,College of Engineering,Southern University of Science and Technology,Shenzhen 518055 Guangdong,ChinaNational Biomedical Imaging Center,Peking University,Beijing 100871,China||School of Life Science,Peking University,Beijing 100871,ChinaDepartment of Biomedical Engineering,College of Engineering,Southern University of Science and Technology,Shenzhen 518055 Guangdong,China||Eastern Institute for Advanced Study,Eastern Institute of Technology,Ningbo,Zhejiang 315200,China||Institute for Biomedical Materials and Devices(IBMD),Faculty of Science,University of Technology Sydney,Sydney,NSW 2007,AustraliaDepartment of Biomedical Engineering,College of Future Technology,Peking University,Beijing 100871,China||National Biomedical Imaging Center,Peking University,Beijing 100871,China||Airy Technologies Co.,Ltd.,Beijing 100086,ChinaDepartment of Biomedical Engineering,College of Engineering,Southern University of Science and Technology,Shenzhen 518055 Guangdong,China||Airy Technologies Co.,Ltd.,Beijing 100086,ChinaDepartment of Biomedical Engineering,College of Future Technology,Peking University,Beijing 100871,China||National Biomedical Imaging Center,Peking University,Beijing 100871,ChinaAiry Technologies Co.,Ltd.,Beijing 100086,ChinaDepartment of Biomedical Engineering,College of Future Technology,Peking University,Beijing 100871,China||Wallace H.Coulter Dept.of Biomedical Engineering,Georgia Institute of Technology and Emory University,Atlanta 30332 GA,USADepartment of Biomedical Engineering,College of Engineering,Southern University of Science and Technology,Shenzhen 518055 Guangdong,China||Eastern Institute for Advanced Study,Eastern Institute of Technology,Ningbo,Zhejiang 315200,China
《光：科学与应用（英文版）》 2024 (006)
1140-1158 / 19
We thank Airy Technologies Co.,Ltd.for providing us with Airy Polar-SIM system to further build up the PAR-SIM system and collect the data.We thank Yiwei Hou's help with the MRA43 post-processing and test on our data.This work was supported by the National Key R&D Program of China(2022YFC3401100)and the National Natural Science Foundation of China(62025501,92150301,62335008).
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