高分辨率大视场线扫描共焦显微镜的设计与研制  

Design of high‐resolution wide field of view confocal line scanning laser microscopy

作　　者：孔文[1,2] 郎婷婷[2] 高峰[1] 樊金宇 史国华[1] Wen Kong Tingting Lang Feng Gao Jinyu Fan Guohua Shi(Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, China College of Optical and Electronic Technology, China Jiliang University, Hangzhou 310018, China)

机构地区：[1]中国科学院苏州生物医学工程技术研究所,江苏苏州215163 [2]中国计量大学光学与电子科技学院,杭州310018

出　　处：《光电工程》2017年第6期616-620,共5页Opto-Electronic Engineering

基　　金：国家自然科学基金(61675226;61378090);江苏省杰出青年基金(BK20060010);中国科学院前沿科学重点研究项目(QYZDB-SSW-JSC03);科技部重大科学仪器设备开发专项(2016YFF0102000)

摘　　要：商用共聚焦显微镜使用二维光点扫描对样品成像,成像帧频被限制在30 Hz以内,扫描速度大多在10帧/秒(f/s)左右。为了提高共聚焦系统成像速度,满足生物细胞在体成像的需求,本文使用线光束对样品进行一维扫描照明,成像速度大大提高,同时根据共聚焦成像原理,在线阵CCD前使用狭缝滤除非聚焦平面杂散光以提高成像质量。实验表明,系统光学放大倍率为55倍,横向分辨率高于2.2μm,当线阵CCD以28 k Hz行频扫描成像时,帧频可达50 f/s,通过对动植物细胞成像证明,本系统可用于生物细胞的在体成像。Commercial confocal microscopy usually utilizes two-dimensional galvanometer to scan specimen, the frame rate is limited within 30 f/s, and commercial confocal microscopy is 10 f/s or lower. In order to improve the imaging speed of confocal system, and meet the needs of real-time observation of in vivo imaging, in the high-speed confocal line scanning laser microscopy, the sample is illuminated by one-dimensional scanning laser beam. The imaging speed is greatly improved. At the same time, a slit is placed before the linear array CCD to filter out to the non-focused plane stray light and improve the image quality. Experiments show that the optical magnification is 55, horizontal resolution is higher than 2.2 μm, and the frame rate is up to 50 frame per second when the linear CCD scans with 28 k Hz horizontal frequency and 512 pixels 2048 pixels image resolution, image experiment of plant and animal cells prove that this imaging system could be used in ex vivo or in vivo cell image.

关 键 词：成像系统 线扫描 共聚焦 高帧频成像 

分 类 号：TN16[电子电信—物理电子学]