用于双光子激发荧光寿命显微成像的高重复频率皮秒扫描相机
A High Repetition Rate Picosecond Streak Camera for Two-Photon Excited Fluorescence Lifetime Microscopic Imaging
摘要
建立了一台基于新研制的高重复频率皮秒扫描相机的双光子激发荧光寿命显微成像系统,重点介绍所研制的高重复频率皮秒扫描相机。为了在高时间分辨力的同时扩大时间测量范围,实现大面积两维空间高时间分辨取样测量,从而提高采样速率和更有效地发挥扫描相机的作用,设计和研制了一种大面积、高时间分辨力扫描变像管和一种重复频率高达1MHz的斜坡电压扫描电路。基于上述关键部件所研制的扫描相机具有重复频率高、扫描速度可调、时间分辨力高、工作面积大、非线性低、触发晃动小等优点。用钛宝石飞秒激光器作为激光脉冲源,通过脉冲提取器将76 MHz的高重复频率降低为1 MHz,采用可调延时器和标准具对扫描相机的时间分辨力、扫描速度和非线性进行标定。该系统的时间分辨力达到6.5 ps,非线性为2.60%,可测量的时间范围从十几皮秒到几十纳秒。测量了若丹明6G和香豆素314两种标准荧光染料的荧光寿命,取得了与参考文献一致的实验结果。
Abstract
A two-photon excited fluorescence lifetime microscopic imaging system based on a newly-developed high repetition rate picosecond streak camera is presented. The high repetition rate picosecond streak camera is the emphasis for introduction. In order to increase the measurable sweeping range and maintain high temporal resolution as well, and in order to realize high temporally resolved sampling measurement in two dimensions to improve sampling speed and exploit effectively the advantages of the streak camera, authors specially designed and developed a streak image tube with large area and high temporal resolution and a ramp voltage streak circuit with repetition rate up to 1 MHz. The streak camera based on the key components above mentioned is characterized with high repetition rate, tunable sweep rate, high temporal resolution, large effective area of work, low nonlinearity and small triggering jitter. A Ti:sapphire laser provides mode-locked, ultrafast femtosecond pulses with a fundamental frequency of 76 MHz which is reduced to 1 MHz with a pulse picker. The temporal resolution, sweep rate and nonlinearity are calibrated with a tunable delay generator and etalons. Temporal resolution of the system is 6.5 ps, nonlinearity is 2.60%, and the measurable time range is from ten picoseconds to tens of nanoseconds. Streak images of Rhodamine 6G and Coumarin 314 are obtained and the results agree well with those reported in the literatures.
中图分类号:TB872; O433
所属栏目:成像系统
基金项目:国家自然科学基金(60138010, 60408011)资助课题。
收稿日期:2005-05-23
修改稿日期:2005-08-15
网络出版日期:--
作者单位 点击查看
屈军乐:深圳大学光电子学研究所,深圳 518060
林子扬:深圳大学光电子学研究所,深圳 518060
陈丹妮:深圳大学光电子学研究所,深圳 518060
胡涛:深圳大学光电子学研究所,深圳 518060
郭宝平:深圳大学光电子学研究所,深圳 518060
牛憨笨:深圳大学光电子学研究所,深圳 518060
联系人作者:刘立新(lilyliulixin@163.com)
备注:刘立新(1977~),女,河北保定人,中国科学院西安光学精密机械研究所博士研究生,主要从事生物医学光子学的研究。
【1】D. Elson, S. Webb, J. Siegel et al.. Biomedical applications of fluorescence lifetime imaging[J]. Optics & Photonics News, 2002, 13(11): 26~32
【2】K. W. Eliceiri, Ching-Hua Fan, G. E. Lyons et al.. Analysis of histology specimens using lifetime multiphoton microscopy[J]. J. Bio. Opt., 2003, 8(3): 376~380
【4】K. Dowling, S. C. W. Hyde, J. C. Dainty et al.. 2-D fluorescence lifetime imaging using a time-gated image intensifier[J]. Opt. Commun., 1997, 135(1~3): 27~31
【5】W. Becker, A. Bergmann, K. Knig et al.. Picosecond fluorescence lifetime microscopy by TCSPC imaging[C]. Proc. SPIE, 2001, 4262: 414~419
【6】W. Becker, A. Bergmann, C. Biskup et al.. High resolution TCSPC lifetime imaging[C]. Proc. SPIE, 2003, 4963: 175~184
【7】R. V. Krishnan, H. Saitoh, H. Terada et al.. Development of a multiphoton fluorescence lifetime imaging microscopy system using a streak camera [J]. Rev. Scientific Instruments, 2003, 74(5): 2714~2721
【10】Qu Junle, Liu Lixin, Guo Baoping et al.. Development of a multispectral multiphoton fluorescence lifetime imaging microscopy system using a streak camera[C]. Proc. SPIE, 2005, 5630: 510~516
【11】Guo Baoping, Tian Jindong, Qu Junle et al.. Research on the time resolved multi-spectral detection technology[J]. China Basic Science, 2004, (6): 11~14 (in Chinese)
郭宝平,田劲东,屈军乐 等. 时间分辩多光谱技术研究[J]. 中国基础科学, 2004, (6): 11~14
【13】Liu Yonggang, Peng Xiaoshi, Tang Xiaoqing. Measuring sweep speed and sweep nonlinearity of optic streak camera[J]. Nuclear Electronics & Detection Technology, 2004, 24(2): 218~220 (in Chinese)
刘永刚,彭晓世,汤小青. 光学条纹相机扫描速度和非线性的测试[J]. 核电子学与探测技术, 2004, 24(2): 218~220
【15】M. J. Cole, J. Siegel, S. E. D. Webb et al.. Time-domain whole-field fluorescence lifetime imaging with optical sectioning[J]. J. Microsc., 2001, 203(3): 246~257
【16】R. Cubeddu, D. Comelli, C. D′Andrea et al.. Time-resolved fluorescence imaging in biology and medicine[J]. Appl. Phys., 2002, 35(9): R61~R67
【17】H. Zeng, M. Petek, M. T. Zorman et al.. Integrated endoscopy system for simultaneous imaging and spectroscopy for early lung cancer detection[J]. Opt. Lett., 2004, 29(6): 587~589
【18】J. Siegel, D. S. Elson, E. D. Webb et al.. Studying biological tissue with fluorescence lifetime imaing: microscopy, endoscopy, and complex decay profiles[J]. Appl. Opt., 2003, 42(16): 2995~3004
引用该论文
刘立新,屈军乐,林子扬,陈丹妮,胡涛,郭宝平,牛憨笨. A High Repetition Rate Picosecond Streak Camera for Two-Photon Excited Fluorescence Lifetime Microscopic Imaging[J]. Acta Optica Sinica, 2006, 26(3): 373-378
刘立新,屈军乐,林子扬,陈丹妮,胡涛,郭宝平,牛憨笨. 用于双光子激发荧光寿命显微成像的高重复频率皮秒扫描相机[J]. 光学学报, 2006, 26(3): 373-378
被引情况
【1】牛丽红,刘进元,彭文达,牛憨笨,杨勤劳,廖华,周军兰,张劲松. 微通道板选通X射线纳秒分幅相机的研制. 光学学报, 2008, 28(7): 1274-1278
【3】王岩,李恒,齐璟,邵永红,屈军乐,牛憨笨. 基于扫描相机的时域荧光寿命测量及分析. 中国激光, 2011, 38(s1): 108001--1