基于数字微镜器件的快速超分辨晶格结构光照明显微研究  

Digital micromirror device-based fast super-resolution lattice structured light illumination microscopy

作　　者：杨浩智 聂梦娇 马光鹏 曹慧群 林丹樱[1] 屈军乐[1] 于斌[1] Yang Hao-Zhi;Nie Meng-Jiao;Ma Guang-Peng;Cao Hui-Qun;Lin Dan-Ying;Qu Jun-Le;Yu Bin(Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province,College of Physics and Optoelectronic Engineering,Shenzhen University,Shenzhen 518060,China;College of Chemistry and Environmental Engineering,Shenzhen University,Shenzhen 518060,China)

机构地区：[1]深圳大学物理与光电工程学院,光电子器件与系统教育部/广东省重点实验室,深圳518060 [2]深圳大学化学与环境工程学院,深圳518060

出　　处：《物理学报》2024年第9期297-306,共10页Acta Physica Sinica

基　　金：国家重点研发计划(批准号:2022YFF0712500);国家自然科学基金(批准号:62175166,61975131,62235007,62275165,62127819);深圳市科技计划(批准号:JCYJ20200109105411133,JCYJ20220818100202005);深圳市光子学和生物光子学重点实验室(批准号:ZDSYS20210623092006020)资助的课题

摘　　要：超分辨结构光照明显微成像技术(super-resolution structured illumination microscopy,SR-SIM)具有时间分辨率高、光漂白和光毒性低和对荧光探针的要求少等优点,适用于活细胞的长时程超分辨成像.采用二维晶格结构光作为照明光,可以实现更快的成像速度和更低的光毒性,但同时也增加了系统的复杂性.为了解决此问题,本文提出了一种基于数字微镜器件的快速超分辨晶格结构光照明显微成像方法(digital micromirror device-based lattice SIM,DMD-Lattice-SIM),通过同步分时触发DMD和sCMOS相机的方式实现二维正交晶格结构光的产生,且只需要采集5幅相移原始图像即可重构出超分辨图像,相比于传统SR-SIM需要9幅相移原始图像的方法,图像采集时间减少了约44.4%.同时,在基于空域和频域联合的SIM重构算法(joint space and frequency reconstruction method-SIM,JSFR-SIM)的基础上,本文还发展了用于Lattice-SIM的JSFR超分辨图像重构方法(Lattice-JSFR-SIM),先在频域对原始图像进行预滤波处理;然后,在空域对滤波后的图像进行超分辨重构处理.与传统频域图像重构处理对比,该方法在512×512像素数的成像视场下重构时间减少了约55.6%,对于实现活细胞实时超分辨成像具有重要意义和应用价值.Super-resolution structured illumination microscopy(SR-SIM)offers numerous advantages such as high temporal resolution,low photobleaching and phototoxicity,and no special requirements for fluorescent probes.It is particularly suitable for long-term SR imaging of living cells.By using two-dimensional lattice structured light serving as illumination,SR-SIM can achieve faster imaging speed and reduce phototoxicity,however,it is accompanied with system complexity increasing.To address this problem,in this work,we propose a fast SR lattice structured illumination microscopy imaging method based on a digital micromirror device(DMD),called DMD-Lattice-SIM.This method utilizes a DMD and synchronous time-sharing triggering with sCMOS to generate two-dimensional orthogonal lattice structured light.The proposed method only requires the collection of five phase-shifted raw images for SR image reconstruction,reducing the acquisition time by approximately 44.4%compared with the traditional SR-SIM method that requires nine phase-shifted raw images.In this work,we also introduce a rapid SR image reconstruction method called Lattice-JSFR-SIM,which combines the advantages of joint space and frequency reconstruction(JSFR)-SIM and Lattice-SIM.The raw images are pre-filtered in the frequency domain and then undergo SR reconstruction in the spatial domain.This approach reduces reconstruction time by approximately 55.6%compared with traditional frequency domain image reconstruction processing,within an imaging field of view of 512 pixels×512 pixels.The feasibility of the proposed method is demonstrated through experiments on cell microtubules and the observation of mitochondrial division and fusion in living cells.The findings presented in this paper hold great significance and application value for enabling real-time SR imaging of living cells.

关 键 词：结构光照明显微术 超分辨成像 晶格结构光照明 空域重构 

分 类 号：TH742[机械工程—光学工程]