双路频分复用荧光共焦显微探测技术应用研究.docVIP

双路频分复用荧光共焦显微探测技术研究 唐平玉 郑继红* 曹剑炜 张运波 蒋妍梦 黄爱琴 周增军 庄松林 （上海理工大学 光电信息与计算机工程学院，上海 200093） 摘要 文章报道了结合频分复用技术来提高荧光共焦生物显微探测系统探测能力的实现原理和方法。通过对双路激发光信号的载频调制，将激发光聚焦到生物样品上产生荧光信号，再通过傅立叶变换、滤波和解调制过程，最后将两路荧光信号强度随时间变化曲线进行还原。实验搭建了紫外波段激励光源的双频复用荧光共焦显微成像系统，并成功探测到了老鼠神经海马细胞样品发出的双点荧光信号。频分复用共焦显微成像系统能够实现多通道，实时，快速探测生物细胞荧光信号，并具有较高的时间和空间分辨率，为对生物细胞特别是活体细胞的实时探测提供了一种高效的方法和手段。 关键词 共焦显微；频分复用；荧光；细胞探测 中图分类号 Q336 Q438 OCIS 180.1790? ? 120.0120 110.0110? 110.0180 文献标识码 A Two-channel Frequencies Division Multiplexed Fluorescence Confocal Microscopy Tang Pingyu, Zheng Jihong*, Cao Jianwei, Zhang Yunbo, Jiang Yanmeng, Huang Aiqing, Zhou Zengjun, Zhuang Songlin (School of Optical-electrical and Computer Engineering, University of Shanghai for Science and Technology, Shanghai, 200093, China) Abstract This paper presents a setup of confocal fluorescence microscopy system combined with frequencies division multiplexing so that the detection ability of the system is enhanced. In the frequency divided multiplexed confocal fluorescence microscopy (FDMCFM) system proposed, fluorescence is excited on the sample (rat neural cell) by two beams of light modulated individually in two arms of the system. Fourier transform, filtration and demodulation are then carried out to restore the fluorescence intensity signal. The FDMCFM system is experimentally conducted, through which the target cells are detected successfully. The experimental results show that the FDMCFM system is convenient for both multiple-channels and real-time detection of biological cell’s fluorescence signal. Moreover, it maintains the high spatial and temporal resolution of confocal microscopy. These characteristics of the system make it an effective solution to the dynamic studies of living biological cells. Keywords confocal microscopy; frequency division multiplexing; fluorescence microscopy; cell detection 1 引 言 激光扫描共焦显微镜(Laser Scanning Confocal Microscope，LSCM) 2 基本原理 共焦显微术由Marvin Minsky提出并于1957 年申请了载物台扫描共焦光学显微镜美国国家专利[4]。在此共焦成像系统