High-Speed Line Scanning Confocal Laser Ophthalmoscope

He Yi, Shi Guohua, Lu JingLi Hao, Zhang Yudong
2012 Acta Optica Sinica  
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 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 2.2 μm,当线阵 CCD 以 28 kHz 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。 关键词:成像系统;线扫描;共聚焦;高帧频成像 中图分类号:TN16 文献标志码:A Abstract: Commercial confocal microscopy usually utilizes two-dimensional galvanometer to scan specimen, the frame rate is
more » ... 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 kHz 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.
doi:10.3788/aos201232.0117001 fatcat:5ybwg3exo5g4vfdkdqtnvdh7ba