摘要
亚细胞器是细胞的重要组成单位,其形态结构与动力学特性直接反映了细胞的生理状态。21世纪初新兴的结构光照明显微技术、受激发射损耗显微技术和单分子定位成像技术等超分辨显微成像技术,巧妙地绕过了光学衍射极限对成像分辨率的限制,目前已被广泛应用于活细胞亚细胞器精细结构的观察及其动力学过程的监测上。本文首先介绍了上述三种超分辨显微成像技术的基本原理和特点,然后介绍了活细胞中细胞核、细胞骨架、线粒体、内质网等亚细胞器的超分辨精细结构和动力学特性,最后讨论了亚细胞器超分辨精细结构成像与机器学习、图像处理相结合的发展潜力。
Significance It is well-known that subcellular organelles are essential components of cells.Their morphological structures and dynamic characteristics directly reflect the physiological state of cells.Scientists have paid significant attention to the observation and analysis of the fine structures of subcellular organelles in living specimens.The emerging super-resolution microscopy(SRM)techniques in the early 21st century,such as structured illumination microscopy(SIM),stimulated emission depletion(STED),and single-molecule localization microscopy(SMLM),skillfully bypass the limitation of the optical diffraction limit and effectively retain the advantages of optical microscopy.SRM techniques have been widely used in monitoring subcellular organelles in living cells.Progress This article systematically elaborates and analyzes the super-resolution structure characteristics of subcellular organelles in living cells.First,it briefly introduces the basic principles and fundamental characteristics of the three kinds of SRM techniques,i.e.,STED,SIM,and SMLM,and expounds their development status.Second,the superresolution fine structures and dynamic characteristics of subcellular organelles,such as the nucleus,cytoskeleton,mitochondrion,and endoplasmic reticulum(ER),are presented.In 2016,Chagin et al.quantitatively measured and analyzed replication foci(RF)in mammalian cells using threedimensional(3D)SIM.Mitchell-Jordan et al.(2012)directly imaged histone protein H3in mammalian cells using STED to show the chromatin domain characteristics at the scale of 40--70nm.Wombacher et al.and Lukinavicˇius et al.also employed STORM to observe the distribution of histone protein H2Bin living HeLa and U2OS cells,respectively.Pelicci et al.(2020)imaged nuclear Lamin-A in intact nuclei of living cells through SPLIT-STED.Otsuka et al.(2016)captured images of different steps involved in assembling the NPC in a human cell.Lu et al.and Zhao et al.also realized the NPC super-resolution fluorescence imaging using different methods.Gustafs
作者
王冠晨
陈同生
Wang Guanchen;Chen Tongsheng(Key Laboratory of Laser Life Science,Ministry of Education,College of Biophotonics,South China Normal University,Guangzhou 510631,Guangdong,China;Guangdong Key Laboratory of Laser Life Science,College of Biophotonics,South China Normal University,Guangzhou 510631,Guangdong,China;SCNU Qingyuan Institute of Science and Technology Innovation Co.,Ltd.,Qingyuan 511517,Guangdong,China)
出处
《中国激光》
EI
CAS
CSCD
北大核心
2022年第20期86-94,共9页
Chinese Journal of Lasers
基金
国家自然科学基金(61875056,62135003)
广州市科技计划项目(2019B090905005)。
关键词
生物光学
超分辨显微术
荧光显微镜
亚细胞器精细结构
机器学习
bio-optics
super-resolution microscopy
fluorescence microscope
fine structures of subcellular organelles
machine learning
