基于结构照明的超分辨荧光显微成像重建算法  

Super-Resolution Fluorescence Microscopy Image Reconstruction Algorithm Based on Structured Illumination

作　　者：刘智 罗泽伟 王正印 涂壮 庄正飞 陈同生 Liu Zhi;Luo Zewei;Wang Zhengyin;Tu Zhuang;Zhuang Zhengfei;Chen Tongsheng(Key Laboratory of Laser Life Science,Ministry of Education,College of Biophotonics,South China Normal University,Guangzhou,Guangdong 510631,China;Guangdong Key Laboratory of Laser Life Science,College of Biophotonics,South China Normal University,Guangzhou,Guangdong 510631,China)

机构地区：[1]华南师范大学生物光子学研究院,教育部激光生命科学重点实验室,广东广州510631 [2]华南师范大学生物光子学研究院,广东省激光生命科学重点实验室,广东广州510631

出　　处：《中国激光》2021年第3期85-97,共13页Chinese Journal of Lasers

基　　金：国家自然科学基金(61875056);国家重大科研仪器研制项目(61527825);广州市科技计划项目(2019050001)。

摘　　要：由于具有低光毒性、高速宽视场以及多通道三维超分辨成像能力,超分辨结构照明显微术(SR-SIM)特别适合用于活细胞中动态精细结构的实时检测研究。超分辨结构照明显微图像重建算法(SIM-RA)对SR-SIM的成像质量具有决定性影响。本文首先简要介绍了超分辨显微术的发展现状,阐述了研究SR-SIM图像重建算法的必要性;然后介绍了SR-SIM的成像原理,并重点介绍了SR-SIM图像重建算法,包括SR-SIM中频繁使用的去卷积重建算法、SR-SIM校准与重建过程中参数值获取的算法,以及目前发展的超分辨结构照明显微图像重建算法,并介绍了SR-SIM工具箱;最后总结了当前发展超分辨结构照明显微图像重建算法需解决的5个问题。Significance Super-resolution microscopy overcomes the 200 nm spatial resolution limitation of traditional optical microscopy and has been widely used to image subcellular structures in living cells.Super-resolution microscopy mainly includes stimulated emission depletion(STED) microscopy,structured illumination microscopy(SIM),and single-molecule localization microscopy(SMLM).Super-resolution structured illumination microscopy(SR-SIM) is the most widely used super-resolution microscopy technique.Owing to the low phototoxicity,high-speed wide field of view,and multichannel three-dimensional super-resolution imaging capabilities,SR-SIM is especially suitable for real-time detection of dynamic fine structures in living cells.SR-SIM has high-speed wide-field imaging capabilities that enable rapid dynamic measurement of living cells.Compared with STED and SMLM microscopy imaging under a light intensity of~1000 W/cm2,SR-SIM allows imaging under a low light intensity of ~10 W/cm2,which meets the requirement of long-term imaging of living cells.At the same time,multicolor and multichannel imaging by SRSIM is beneficial to the study of molecular interactions.The super-resolution structured illumination reconstruction algorithm(SIM-RA) is the key to realizing fast,long-term,and nondestructive super-resolution imaging of SR-SIM.SR-SIM uses structured light in different directions and phases to excite the fluorescent sample for acquiring multiple original images with illumination patterns and then reconstructs a single super-resolution image through mathematical calculations.Owing to the imperfection of the experiment and the inevitable uncertainty in the measurement of experimental parameters,the estimation of the reconstruction parameters results in false signal and noise,which reduces the resolution of the SR-SIM image and introduces artifacts that can be misinterpreted as biologically relevant features.An excellent SIM-RA can improve the accuracy of reconstruction parameters and filter out noise to reduce artifacts and

关 键 词：生物光子学 光学成像 超分辨显微术 结构照明显微术 图像重建算法 荧光 多帧重建 

分 类 号：O436[机械工程—光学工程]