快速宽场三维显微技术研究进展  

Advances in Rapid Three-Dimensional Wide Field Microscopy

作　　者：任婧荣 傅相达 王孟瑞 赵天宇[1,2] 汪召军 冯坤 梁言生 王少伟 雷铭 Ren Jingrong;Fu Xiangda;Wang Mengrui;Zhao Tianyu;Wang Zhaojun;Feng Kun;Liang Yansheng;Wang Shaowei;Lei Ming(MOE Key Laboratory for Nonequilibrium Synthesis and Modulation of Condensed Matter,Xi an 710049,Shaanxi,China;School of Physics,Xi an Jiaotong University,Xi an 710049,Shaanxi,China)

机构地区：[1]教育部物质非平衡合成与调控重点实验室,陕西西安710049 [2]西安交通大学物理学院,陕西西安710049

出　　处：《中国激光》2023年第3期49-64,共16页Chinese Journal of Lasers

基　　金：国家重点研发计划(2022YFF0712500);国家自然科学基金(62135003,62005208,62205267,62205265);陕西省创新能力支撑计划(2021TD57);陕西省自然科学基础研究计划(2022JZ34,2020JQ072,2022JQ069,2022JM321)。

摘　　要：激光扫描显微镜通过扫描高度汇聚的激光焦点可以获得样品的三维图像,而激光扫描显微镜时间分辨率低、光毒性大的缺点限制了其在活体快速三维成像等领域中的应用。近年来具有三维成像能力的宽场显微镜技术逐渐成为三维成像领域的研究热点。聚焦形貌恢复技术、结构光照明显微技术以及深度学习辅助三维成像是三种基于宽场成像的快速三维成像技术,通过硬件提升和软件辅助的方式,提高了宽场显微镜的三维成像能力。分别介绍了它们的原理、优缺点、最新的研究进展与应用,最后对宽场三维显微技术的未来发展进行了总结与展望。Significance Threedimensional(3D)imaging is an important research direction in microscopy and has been applied to many fields such as biomedicine and engineering science.Typical 3D microscopy techniques,such as laser confocal microscopy and multiphoton microscopy,are based on laser point scanning geometry,and the imaging speed is limited by the scanning speed;therefore,biological samples are likely to be damaged under long scanning durations and highintensity illumination.Recently,widefield microscopy with 3D imaging capability has received significant attention.Widefield microscopy can yield complete twodimensional imaging simultaneously and affords temporal resolutions higher than spot scanning by two to three orders of magnitude.Additionally,widefield imaging offers highquality grayscale images and fewer samples to be damaged,thus rendering it suitable for the realtime observation of living samples.However,conventional widefield microscopy suffers from defocused backgrounds and low axial resolutions.Owing to the rapid development of computer science and optical technology,various algorithms and techniques for processing widefield images have been proposed to improve their axial resolutions,thus providing more possibilities for 3D imaging.We focus on three types of rapid 3D widefield microscopy techniques,i.e.,shape of focus(SFF),structural illumination microscopy(SIM),and deep learningassisted 3D imaging.The SFF technique enables the extraction of focal plane information by processing a series of image stacks and reconstructing the 3D morphology of samples without requiring specific hardware.In SIM,samples are illuminated by phaseshifted light fields with high spatial frequencies,images are captured using a CCD camera,and the in-and outfocus information can be effectively separated using decoding algorithms.Deep learning models can learn the mapping relationship between different types of images from a large amount of data,such as the conversion between widefield images and confocal images;this is a simple met

关 键 词：生物光学 宽场显微镜 三维成像 聚焦形貌恢复 结构光照明显微镜 深度学习 

分 类 号：O436[机械工程—光学工程]