激光与光电子学进展, 2020, 57 (24): 240001, 网络出版: 2020-11-20
高速超分辨结构光照明显微的关键技术及应用 下载: 4142次封面文章特邀综述
High-Speed Structured Illumination Microscopy and Its Applications
显微 荧光显微镜 超分辨显微镜 结构光照明显微镜 硬件设计方法 图像重建算法 microscopy fluorescence microscopy super-resolution microscopy structured illumination microscopy hardware design image reconstruction algorithm
摘要
光学显微成像技术无论是在临床诊疗还是在基础科学研究上都发挥着重要的作用。伴随着新型荧光探针、光学控制、探测器件的不断发展,超分辨光学显微技术突破了传统光学衍射极限的限制,为现代生物医学研究提供了新的工具。在超分辨显微成像技术中,结构光照明显微镜(SIM)通过空间编码的结构光照明样品,将样品部分超出衍射极限的高频信息调制到低频中,从而通过光学系统实现超分辨成像。SIM具有成像速度快,光漂白和光毒性弱以及对荧光染料的非特异性需求等优点,被广泛应用于活细胞超分辨光学显微成像。本文回顾了SIM技术的重要原理与技术进步,重点介绍了SIM硬件设计与图像重构算法中关键的实验要点与技术难点,列举了现阶段SIM在生物成像中的部分应用,探讨了SIM未来的发展方向。期望本文能为SIM的设计和使用者提供一定的指导。
Abstract
Optical microscopy performs an increasingly important role in clinical diagnosis and basic scientific research. With the development of novel fluorescence probes, light controllers, and detectors, super-resolution optical microscopy breaks through the diffraction limit and provides new tools for modern biomedical research. Among these techniques, structured illumination microscope (SIM) achieves super-resolution by using spatially coded structured illumination which down modulates spatial frequencies beyond the cutoff into the pass band of the microscope. SIM shows lower photo bleaching and phototoxicity, higher imaging speed, and no special requirements for fluorescent probes, which has significant advantages in application to live-cell biomedical research. In this paper, the important principles and technological progress during the development of SIM are firstly reviewed. Then we focus on the key experimental techniques and difficulties in hardware design and image reconstruction of SIM. Finally, the several applications in biological imaging are listed. It is expected that this review can provide guidance for designing and using SIM.
赵天宇, 汪召军, 冯坤, 梁言生, 何旻儒, 云雪, 雷铭. 高速超分辨结构光照明显微的关键技术及应用[J]. 激光与光电子学进展, 2020, 57(24): 240001. Tianyu Zhao, Zhaojun Wang, Kun Feng, Yansheng Liang, Minru He, Xue Yun, Ming Lei. High-Speed Structured Illumination Microscopy and Its Applications[J]. Laser & Optoelectronics Progress, 2020, 57(24): 240001.