单分子定位超分辨成像技术进展及应用  

Single-Molecule Localization Super-Resolution Microscopy and Its Applications

作　　者：杨建宇 董浩 邢福临 胡芬 潘雷霆 许京军 Yang Jianyu;Dong Hao;Xing Fulin;Hu Fen;Pan Leiting;Xu Jingjun(Key Laboratory of Weak-Light Nonlinear Photonics,Ministry of Education,School of Physics and TEDA Institute of Applied Physics,Nankai University,Tianjin300071,China;State Key Laboratory of Medicinal Chemical Biology,College of Life Sciences,Frontiers Science Center for Cell Responses,Nankai University,Tianjin300071,China)

机构地区：[1]弱光非线性光子学教育部重点实验室,南开大学物理科学学院,泰达应用物理研究院,天津300071 [2]药物化学生物学国家重点实验室,南开大学生命科学学院,细胞应答交叉科学中心,天津300071

出　　处：《激光与光电子学进展》2021年第12期1-35,共35页Laser & Optoelectronics Progress

基　　金：广东省基础与应用基础研究重大项目(2020B0301030009);国家自然科学基金(11874231,31801134,31870843);中国博士后科学基金(2020M680032);南开大学中央高校基本科研业务费专项资金(2122019446);高等学校学科创新引智计划(B07013)。

摘　　要：21世纪初诞生的超分辨光学成像技术凭借纳米级空间分辨率、低损制样等优点,迅速发展成为生命科学研究中不可或缺的技术手段。其中单分子定位超分辨成像(SMLM)技术更是由于其成像原理易懂、空间分辨率极高等特点,一直受到科研工作者的青睐,不断取得重要的技术和应用进展。首先回顾了SMLM的工作原理,讨论了其光路搭建、图像重建、漂移校正等关键技术问题。介绍了两类代表性SMLM技术。列举了多种多色SMLM方法,并分析了各自的优缺点。介绍了SMLM成像参数的改进研究,包括横/纵分辨率的提高、成像视野和深度的改善。介绍了SMLM和深度学习,SMLM和电镜等成像手段结合的关联成像研究进展。讨论了SMLM数据提取与分析方法。最后列举了SMLM在细胞生物学中的重要应用,并展望了SMLM未来的发展方向。期望该综述能为SMLM工作者提供有益的启发和参考,推进SMLM在生命科学研究中的深入应用。Super-resolution microscopy invented at the beginning of the 21 st century has rapidly become a indispensable method in life science research owing to their nanoscale spatial resolution,low damage of sample preparation,and so on.Among a variety of super-resolution imaging techniques,single-molecule localization superresolution microscopy(SMLM)with straightforward principle and outstanding spatial resolution gains more and more attention from researchers,thereby continuously making significant progress on the techniques and applications.Firstly,this paper reviewed the principle of SMLM and discussed some technical problems including the optical path building,the image reconstruction,and the drift correction.Two types of representative SMLMs were indroduced and discussed.Then,diversified multi-color SMLMs were introduced and their advantages and disadvantages were analyzed.The improvement of imaging parameters including the lateral/axial spatial resolution,imaging field,and imaging depth of SMLM was subsequently discussed.The research progress of correlated imaging of the SMLM combined with deep learning and SMLM combined with the electron microscope was further introduced.Moreover,the extraction and analysis methods of SMLM data were discussed.Finally,some important applications of SMLM in cell biology were listed and the development prospects of SMLM were discussed.We hope the present review could be a useful reference for the SMLM users and provides novel insights for them,thus promoting the in-depth applications of SMLM in life science research.

关 键 词：显微 荧光显微成像 超分辨成像 单分子定位 光学衍射极限 图像重建 

分 类 号：O436[机械工程—光学工程]