结构光照明显微镜(Structured Illumination Microscopy,SIM)通过结构化照明在频率域以空间混频的方式将物体高频信息载入光学系统的探测通带内实现突破衍射极限的超分辨光学显微成像。SIM凭借其较低的激发光强、对荧光染料的非特异性需求以及快速的宽场成像优势已成为活细胞超分辨光学显微成像方面应用最多的技术。本文系统回顾了SIM的技术进展, 对SIM的基本原理与实现方法进了详细的分析, 重点介绍了本课题组研发的基于光谱分辨的单光子激发超分辨显微镜和结合自适应光学的双光子激发超分辨显微镜这两种最新的SIM技术, 最后简要讨论了SIM技术在生物成像中的应用及未来发展方向。

Structured illumination microscopy(SIM) is capable of providing super-resolution imaging. It breaks the diffraction limit by moving the high-frequency information of objects into the detectable frequency band of the optical imaging system via frequency mixing. Due to its attractive advantages, such as low intensity illumination, independence of particular fluorescent dyes, and rapid wide-field imaging capability, SIM has become the most popular technique for super-resolution imaging of living cells. This paper first systematically summarizes advances in the development of SIM and introduces corresponding principles at the same time. Then, two novel techniques of SIM developed by our group, including the single-photon excited super-resolution microscopy based on spectral unmixing and the two-photon excited super-resolution microscopy combined with adaptive optics, are particularly introduced in detail. At last, the recent applications and future directions of SIM in biological imaging are briefly discussed.