基于激光干涉的结构光照明超分辨荧光显微镜系统  

Super-Resolution Fluorescence Microscopy System by Structured Light Illumination Based on Laser Interference

作　　者：文刚[1] 李思黾[1] 杨西斌[1] 王林波[1] 梁永[1] 金鑫[1] 朱茜[1] 李辉[1] 

机构地区：[1]中国科学院苏州生物医学工程技术研究所,江苏省医用光学重点实验室,江苏苏州215163

出　　处：《光学学报》2017年第3期25-35,共11页Acta Optica Sinica

基　　金：国家自然科学基金(11504409,61405238,61475185);江苏省自然科学基金(BK20150357,BK20141206);中国科学院科研装备研制项目(YZ201443)

摘　　要：结构光照明荧光显微术(SIM)是一种可突破阿贝衍射极限的宽场显微成像技术,因其非侵入、成像速度快及光损伤小等优点在生物医学研究中具有广泛的应用前景。从结构光照明显微成像系统基本原理出发,分析了超分辨图像重构算法原理、重构图像中伪影来源及优化方法;结合研制的线性/非线性结构光照明显微镜,详细讨论了基于激光干涉的SIM成像系统光机结构。重点讨论了系统的同步时序设计和光路中的几个关键技术问题。设计对比实验验证了自主开发的SIM重构算法的可靠性,并基于研制的线性SIM系统开展细胞骨架的成像实验。最后,对SIM技术在生物上的发展和应用提出展望。Structured illumination fluorescence microscopy（SIM）is a wide field microscopy imaging technique which can circumvent the Abbe diffraction limit.It has various potential applications in the biomedical research due to its advantages of non-invasivity,fast imaging speed and low photon damage and so on.Based on the basic principle of structured illumination microscopy imaging system,the super-resolution image reconstruction algorithm,the sources of artifacts in reconstructed image and optimization methods are analyzed.The SIM imaging system based on laser interference is discussed combining custom built linear/non-linear structured illumination microscope.Design of the control system for hardware synchronization,and few key questions during optical alignment are discussed in detail.Imaging of cell skeleton is done with the linear SIM microscopy system and the comparative tests verify the reliability of the independent-developed image reconstruction algorithm.At last,the future improvement and application in biology of the SIM technique are discussed.

关 键 词：显微 超分辨 荧光显微镜 结构光照明 激光干涉 

分 类 号：O436[机械工程—光学工程]