基于大视场结构光照明的三维层析显微成像技术  

Three dimensional tomographic microscopy based on large-field optical sectioning structured illumination

作　　者：温同强 陈肖霏 温凯[2,3,4] 郜鹏 WEN Tongqiang;CHEN Xiaofei;WEN Kai;GAO Peng(School of Mechanical and Electronic Engineering,Luoyang Institute of Science and Technology,Luoyang 471000,Henan,China;School of Physics,Xi'dian University,Xi'an 710071,Shaanxi,China;Key Laboratory of Optoelectronic Perception of Complex Environment,Ministry of Education,Xi'an 710071,Shaanxi,China;Engineering Research Center of Information Nanomaterials,Universities of Shaanxi Province,Xi'an 710071,Shaanxi,China)

机构地区：[1]洛阳职业技术学院机电工程学院,河南洛阳471000 [2]西安电子科技大学物理学院,陕西西安710071 [3]复杂环境光电感知教育部重点实验室,陕西西安710071 [4]陕西省高校信息纳米材料工程研究中心,陕西西安710071

出　　处：《陕西师范大学学报（自然科学版）》2024年第6期38-47,共10页Journal of Shaanxi Normal University：Natural Science Edition

基　　金：国家重点研发计划(2021YFF0700303);国家自然科学基金(62075177)。

摘　　要：提出一种大视场结构照明光切片显微(large-field optical sectioning structured illumination microscopy,LF-OS-SIM)技术,以实现对厚样品的三维层析显微成像。该技术利用一维光栅投影生成条纹结构光场,并结合空间光调制器(spatial light modulator,SLM)对结构光的频谱进行快速相移。与传统的结构照明光切片显微技术相比,LF-OS-SIM的成像视场(field of view,FOV)提升至原有的4.7倍。同时,利用空间光调制器(spatial light modulator,SLM)进行数字相移,获得了20帧/s的切片速度。利用LF-OS-SIM对硬币、三维分布的荧光小球、生物样品进行了三维层析显微成像,结果发现LS-OS-SIM的成像视场达1030×780μm 2,轴向层析成像精度达4.0±0.39μm。由于其大视场、高分辨率和快速切片速度的优势,LS-OS-SIM有望在工业微器件和生物样品的三维成像中得到广泛应用。This study introduces a technique known as large-field optical sectioning structured illumination microscopy(large-field optical sectioning structured illumination microscopy,LF-OS-SIM)designed for three-dimensional tomographic microscopic imaging of thick specimens.The technique utilizes a one-dimensional grating projection to generate a striped structured light field,in conjunction with a spatial light modulator(spatial light modulator,SLM)for fast phase-shifting of the structured light spectrum.Compared to conventional OS-SIM techniques that are based on SLM/DMD projection,LF-OS-SIM enhances the spatial bandwidth product(spatial bandwidth product,SBP)by 4.7 times.Additionally,digital phase-shifting using the SLM is implemented to achieve a slicing speed of 20 frames per second.The LF-OS-SIM was employed for three-dimensional tomographic microscopic imaging of objects such as coins,three-dimensionally distributed fluorescent beads,and biological specimens.The results indicated that the imaging field of view(FOV)for LS-OS-SIM reached 1030×780μm 3,with an axial tomographic imaging accuracy of 4.0±0.39μm.Given its wide field of view,high resolution,and rapid slicing capabilities,LS-OS-SIM is anticipated to be extensively applied in the three dimensional imaging of both industrial microdevices and biological samples.

关 键 词：结构光照明 三维成像 光学切片 大视场 

分 类 号：O436[机械工程—光学工程]