一种大视野超分辨结构光照明显微成像方法  

Large-field super-resolution microscopic imaging based on structured illumination

作　　者：胡浩 郜鹏 Hu Hao;Gao Peng(School of Marine Science and Technology&Qingdao Research Institute,Northwestern Polytechnical University,Xi′an 710072,China;School of Physics,Xidian University,Xi′an 710126,China)

机构地区：[1]西北工业大学航海学院青岛研究院,西安710072 [2]西安电子科技大学物理学院,西安710126

出　　处：《仪器仪表学报》2023年第5期177-183,共7页Chinese Journal of Scientific Instrument

基　　金：中央高校基本科研业务费专项(31020190QD034)资助。

摘　　要：近年来结构光照明显微术(SIM)在技术发展和应用方面都受到了广泛关注。然而,传统采用空间光调制器(SLM)进行条纹投影的SIM成像视野有限。为满足生物医学研究中高通量显微成像的需求,本文基于SIM开发一种基于激光干涉的大视野超分辨荧光显微成像系统。将用于投射条纹图案的2D光栅和用于控制条纹方向相移的SLM相结合,突破数字投影设备对条纹数量和精细度的限制。开发一种空域重建超分辨图像的算法来提高成像速度。最后,搭建1套结构光照明荧光显微成像系统样机,在20×放大倍数和物镜NA 0.75的条件下,验证了本文方法的有效性,在分辨率提升至1.8倍的同时视野可达1380μm×1035μm,所述空域超分重建算法能够大幅降低计算耗时。During the past decades,the structured illumination microscopy(SIM)has been drawing great attention for both the technique development and applications.However,the conventional SIM that uses a spatial light modulator for fringe projection often has limited imaging field of view.Therefore,we propose a large-field SIM method that combines a 2D grating for fringe pattern projection and an SLM for selecting fringe orientation and performing phase shifting digitally.The proposed SIM technique breaks the bottleneck of fringe number limited by the digital projection devices,while maintaining the advantage of high-speed(digital)phase shifting of conventional SIM.An algorithm of spatial reconstruction is developed to improve the super-resolution imaging speed.Finally,a SIM fluorescence microscopy system is developed.A large field view of 1380μm×1035μm with a 20×/NA 0.75 objective is experimentally demonstrated,and an enhancement of 1.8 times higher resolution is realized.The spatial reconstruction algorithm can greatly reduce the computational time of super-resolution reconstruction.

关 键 词：结构光照明 超分辨 大视野 2D光栅 空域重建 

分 类 号：TP391[自动化与计算机技术—计算机应用技术]