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超分辨显微技术在活细胞中的应用与发展

Super-resolution microscopy applications and development in living cell

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摘要

细胞是生命体的基本单位和功能单位, 对活细胞内部结构及其功能的研究是了解掌握生命本质的基础之一, 因此活细胞的实时观测对生命科学的发展具有重要意义。传统的光学显微技术受衍射极限的限制, 无法观测200 nm以下的生物结构细节。近20年来, 随着超衍射极限光学理论、技术、器件和荧光探针等方面的快速发展, 超分辨显微成像技术已成为应用于生命科学研究的重要手段。然而, 大多数超分辨显微方法或测量耗时长, 或易引起荧光蛋白漂白/细胞损伤, 在活细胞研究中受到极大限制, 已成为超分辨显微领域重点攻关的方向之一。为此, 文中结合作者在快速超分辨显微技术研究的基础上, 介绍了基于单分子成像的光激活定位显微技术和随机光学重构显微技术、基于荧光非线性可饱和光转换的受激发射显微技术以及基于结构光照明的超分辨显微技术, 并探讨了在活细胞成像中的发展应用。最后, 文中展望了超分辨显微成像技术在活细胞成像中的未来发展趋势。

Abstract

Cell is the basic unit and functional unit of living body. The study of the internal structure and function of living cells is one of the foundations of mastering the essence of life. Therefore, the real-time observation of living cells is of great significance for the development of life sciences. Conventional optical microscopy is limited by the diffraction limits and can not observe the details of biological structures below 200 nm. In the past 20 years, with the rapid development of super-diffraction limit optical theory, technology, devices and fluorescent probes, super-resolution microscopy has become an important method for life science research. However, most super-resolution microscopic methods or measurements take a long time, or are likely to cause photobleaching/phototoxicity, and are severely limited in living cell studies. In this paper, based on the study of fast super-resolution microscopy, the photoactivated localization microscopy and stochastic optical reconstruction microscopy were introduced based on single molecule localization microscopy. The stimulated emission depletion microscopy based on fluorescence non-linear saturated light conversion and structured illumination microscopy based on structured light illumination was also introduced. Besides, the development and application of cell imaging were explored. Finally, an outlook of the future development trend of super-resolution microscopy in living cell imaging was provided.

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中图分类号:O439

DOI:10.3788/irla201746.1103002

所属栏目:特约专栏-野超分辨成像技术

基金项目:国家重点研发计划(2017YFF0107003); 天津市应用基础及前沿技术研究计划重点项目(15JCZDJC31600)

收稿日期:2017-10-10

修改稿日期:2017-11-20

网络出版日期:--

作者单位    点击查看

胡春光:天津大学 精密仪器与光电子工程学院, 天津 300072
查日东:天津大学 精密仪器与光电子工程学院, 天津 300072
凌秋雨:天津大学 精密仪器与光电子工程学院, 天津 300072
何程智:天津大学 精密仪器与光电子工程学院, 天津 300072
李奇峰:天津大学 精密仪器与光电子工程学院, 天津 300072
胡晓东:天津大学 精密仪器与光电子工程学院, 天津 300072
胡小唐:天津大学 精密仪器与光电子工程学院, 天津 300072

联系人作者:胡春光(cghu@tju.edu.cn)

备注:胡春光(1981-), 男, 副教授, 博士, 主要从事超分辨显微技术、光学精密测量等方面的研究。

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引用该论文

Hu Chunguang,Zha Ridong,Ling Qiuyu,He Chengzhi,Li Qifeng,Hu Xiaodong,Hu Xiaotang. Super-resolution microscopy applications and development in living cell[J]. Infrared and Laser Engineering, 2017, 46(11): 1103002

胡春光,查日东,凌秋雨,何程智,李奇峰,胡晓东,胡小唐. 超分辨显微技术在活细胞中的应用与发展[J]. 红外与激光工程, 2017, 46(11): 1103002

被引情况

【1】陈丹妮,李亚晖,刘 伟,刘正一. 基于和频效应和环形光的超分辨红外显微成像方法. 红外与激光工程, 2018, 47(8): 804003--1

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