大视场结构光照明显微技术(特邀)  

Large-field Structured Illumination Microscopy(Invited)

作　　者：郜鹏 方翔 温凯[1] 雷云泽 熊子涵 李娇月 刘星[1] 郑娟娟[1] 安莎 GAO Peng;FANG Xiang;WEN Kai;LEI Yunze;XIONG Zihan;LI Jiaoyue;LIU Xing;ZHENG Juanjuan;AN Sha(School of Physics and Optoelectronic Engineering,Xidian University,Xi′an 710071,China)

机构地区：[1]西安电子科技大学物理学院,西安710071

出　　处：《光子学报》2022年第8期126-136,共11页Acta Photonica Sinica

基　　金：中央高校基本科研业务费专项资金(No.QTZX22039);国家重点研发计划(Nos.2022YFE0100700,2021YFF0700300);国家自然科学基金(No.62075177);中国轻工业浓香型白酒固态发酵重点实验室开放基金(No.2019JJ012);陕西省自然科学基金(Nos.2020JM-193,2020JQ-324);瞬态光学与光子技术国家重点实验室开放基金(No.SKLTOP202001)。

摘　　要：结构光照明显微(SIM)具有成像速度快、对样品损伤小,以及对荧光标记物和标记程序无特殊要求等优点,成为研究活细胞结构和动态过程的重要成像手段之一。介绍了一种大视场、双模式(条纹/点阵)SIM显微技术。该技术结合二维光栅投影产生条纹/点阵结构光,空间光调制器选择条纹方向和实现相移,打破传统SIM中条纹数量受数字投影设备像素个数限制的瓶颈,同时保持了传统SIM成像速度快的优势。实验结果表明:该技术在20×/0.75数值孔径物镜下可获得690μm×517μm成像视场和1.8倍空间分辨率提升,其空间带宽积是传统SIM的3倍。该技术有望被应用于生物学、化学和工业等领域。Structured Illumination Microscopy(SIM)has the merits of fast imaging speed,minimal invasion,and no additional requirement on fluorophores and labeling strategies,and hence it is one of the premier imaging tools for studying the structure and dynamic processes of living cells.In this paper,we report a large-filed,dual-modality structured illumination microscopy technique,which is constructed by combining a 2D grating for fringe/lattice projection and a spatial light modulator(SLM)for selecting fringe orientation and phase-shifting.Both stripe SIM(using fringe patterns)and lattice SIM(using lattice patterns)can be performed using the same setup,providing super-resolution images and optically sectioned images.First,a 2D grating is used to project fringe or lattice patterns.The interference of the±1 diffraction orders of the grating in the-x and-y directions yields two groups of fringe patterns,forming 2D grid patterns if superimposed.Two polarizers with their polarization azimuth of 90°and 0°(i.e.,-s and-p polarizations)are respectively placed on the spectra of the±1orders along the-x and-y directions and these polarizations can be maintained during the whole imaging process.Hence,the two groups of fringe patterns along the-x and-y directions are incoherent.Nowadays,advanced manufacturing techniques can fabricate larger-scale gratings with small periods,of which the number is tens of folds of the patterns generated by SLMs or Digital Micromirror Devices(DMDs).Second,an SLM is used to perform phase shifting and select the orientation of fringe illumination.Specifically,phase shifting of the fringe/lattice patterns is performed by phase modulating the+1storders of the generated fringe/lattice field on a plane close to the Fourier plane,where the spectra of the±1 diffraction orders are separate from each other.We mean to set a defocusing distance between the Fourier plane and the SLM plane,so that the broadlyextended defocused spectrum can be well sampled by hundreds of SLM pixels,avoiding the injury of the SLM b

关 键 词：大视场 超分辨 结构光照明显微 荧光样品 点阵照明 

分 类 号：O436[机械工程—光学工程]