激光扫描实时共聚焦显微成像系统设计

李朝辉

doi:doi:10.3788/gzxb20114005.0667

光子学报, 2011, 40 (5): 667, 网络出版: 2011-06-14

激光扫描实时共聚焦显微成像系统设计

Design and Optimization in Constructing An In-vivo Confocal Laser Scanning Microscopy

论文大纲

李朝辉 ^*

作者单位

中国科学院长春光学精密机械与物理研究所，长春 130033

共聚焦显微镜光学扫描系统同步采集控制 Confocal Laser Scanning Microscopy(CLSM) Optical scanner Scanning and data acquiring control

摘要

共聚焦扫描显微镜已成为生物医学和材料科学领域研究中非常有价值的一种工具.本文给出了一种反射型激光扫描共聚焦显微成像系统的系统结构和具体设计.采用多面体转镜进行水平扫描,摆镜进行垂直扫描.利用商品透镜设计了光学扫描中继系统,采用光电倍增管作为激发出的荧光探测器,同时给出了数据采集和扫描同步控制系统的组成与设计.利用CODE V优化光学扫描系统以获得尽可能小的扫描光斑尺寸和较大的视场,并综合考虑了采样频率、扫描速度和探测器对整个系统性能的影响，从而给出了该型共聚焦显微成像系统的相互匹配的设计参量.分析结果表明：共聚焦扫描系统设计合理可行；从光学扫描系统到PMT探测单元的各项技术指标得到优化，满足实时探测的要求；该系统具有适应性强，易升级，低成本的技术特点，同时可达到同类商品的技术性能.

Abstract

Confocal laser scanning microscopy(CLSM) is an invaluable tool for a wide range of investigations in the biological and medical sciences. A type construction of laser con-focal scanning and fluorescent detection system was provided. A polygon mirror and a galvanometer scanner were used to implement x-y scan. The relay optical system was designed based on commercial lens with optimized consideration to achieve small scan spot size, large Field-of-View and high efficiency. Fluorescence was detected by a photomultiplier tube with excellent signal-to-noise ratio. The scan system control and image acquisition were designed with an optimized scan velocity and sample clock. All these measures could reduce the blurring effect in the whole imaging process and improve the resolution. The analysis results show that the design of the CLSM is reasonable and all components achieve an optimized consideration for in-vivo scanning imaging; this type of CLSM are easily accessible and can be upgraded according to optical requirements; the performance is comparable to available commercial products, but is superior in many aspects of cost, flexibility and versatility.

参考文献

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李朝辉. 激光扫描实时共聚焦显微成像系统设计[J]. 光子学报, 2011, 40(5): 667. LI Zhao-hui. Design and Optimization in Constructing An In-vivo Confocal Laser Scanning Microscopy[J]. ACTA PHOTONICA SINICA, 2011, 40(5): 667.

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