Expanding super-resolution imaging versatility in organisms with multi-confocal image scanning microscopy

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外文摘要：Resolving complex three-dimensional(3D)subcellular dynamics noninvasively in live tissues demands imaging tools that balance spatiotemporal resolution,field-of-view and phototoxicity.Image scanning microscopy(ISM),as an advancement of confocal laser scanning microscopy,provides a 2-fold 3D resolution enhancement.Nevertheless,the relatively low imaging speed has been the major obstacle for ISM to be further employed in in vivo imaging of biological tissues.Our proposed solution,multi-confocal image scanning microscopy(MC-ISM),aims to overcome the limitations of existing techniques in terms of spatiotemporal resolution balancing by optimizing pinhole diameter and pitch,eliminating out-of-focus signals,and introducing a frame reduction reconstruction algorithm.The imaging speed is increased by 16 times compared with multifocal structured illumination microscopy.We further propose a single-galvo scan,akin to the Archimedes spiral in spinning disk confocal systems,to ensure a high-speed and high-accuracy scan without the galvanometer's inertial motion.Benefitting from its high photon efficiency,MC-ISM allows continuous imaging of mitochondria dynamics in live cells for 1000 frames without apparent phototoxicity,reaching an imaging depth of 175 μm.Noteworthy,MC-ISM enables the observation of the inner membrane structure of living mitochondria in Arabidopsis hypocotyl for the first time,demonstrating its outstanding performance.

外文关键词：

super-resolution imagingconfocalimage scanning microscopy

作者：

Wei Ren、Meiling Guan、Qianxi Liang、Meiqi Li、Boya Jin、Guangxing Duan、Liya Zhang、Xichuan Ge、Hong Xu、Yiwei Hou、Baoxiang Gao、Sodmergen、Peng Xi

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作者单位：

Department of Biomedical Engineering,College of Future Technology,Peking University,Beijing 100871,China

National Biomedical Imaging Center,Peking University,Beijing 100871,China

Key Laboratory of Computational Optical Imaging Technology,Chinese Academy of Sciences,Beijing 100094,China

School of Life Sciences,Peking University,Beijing 100871,China

Key Laboratory of Analytical Science and Technology of Hebei Province,College of Chemistry and Material Science,Hebei University,Baoding 071002,China

Airy Technologies Co.Ltd.,Beijing 100086,China

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基金：

National Key R&D Program of ChinaNational Natural Science Foundation of ChinaNational Natural Science Foundation of ChinaNational Natural Science Foundation of ChinaNational Natural Science Foundation of ChinaNational Natural Science Foundation of China

项目编号：

2022YFC34011006202550131971376921503016233500862305004

出版年：

2024

DOI：

10.1093/nsr/nwae303

国家科学评论(英文版)

CSTPCD

ISSN：

年,卷(期)：2024.11(9)