| 题名 | 双光子显微成像系统关键技术研究 |
| 作者 | 娄艳阳 |
| 学位类别 | 硕士 |
| 答辩日期 | 2015-11 |
| 授予单位 | 中国科学院大学 |
| 导师 | 熊大曦 |
| 关键词 | 双光子 群延迟色散 色散补偿 光束整形 误差扩散法 |
| 其他题名 | Research on Key Technologies in Two-photon Microscopy Imaging System |
| 学位专业 | 机械电子工程 |
| 中文摘要 | 双光子显微成像技术是结合了激光扫描共聚焦技术和双光子激发技术的一种新型光学成像技术。由于其成像深度深、光漂白光毒性小，因此特别适合活细胞、活胚胎及活组织的延时成像，并已经在生命科学领域得到了广泛的应用。本文设计并搭建了一套双光子显微成像原理系统，完成了系统的群延迟色散的测量与补偿，并利用此系统进行了双光子成像。另外，本文在多焦点共聚焦快速扫描方面做了一些研究工作。 首先设计了一套倒置双光子成像系统，分析了系统中的共轭关系和物像关系。对系统元件的选型做了详细的解释，最后对双光子系统的控制流程和成像软件的实现功能做了介绍。 随后，对双光子显微成像系统的群延迟色散进行了校正,提高了双光子激发效率。采用自相关仪测量的方法在自行搭建的双光子系统光路的四个位置测量飞秒激光的脉冲展宽情况，测量样品位置5个波长下最优的群延迟色散补偿值，由此拟合得到自搭建双光子系统的全波段群延迟色散补偿曲线。实验结果表明在应用此群延迟色散补偿曲线后样品位置的脉冲宽度平均减小95 fs，在两个典型激发波长（750 nm和900 nm）生物样品的荧光强度分别提高了42.7%和76.8%。结论为双光子激发效率与飞秒激光的脉冲宽度成线性反比关系。 接下来，对强激光进行了单平顶、多平顶的空间光束整形。采用数字微镜阵列和误差扩散法，对单平顶和多平顶的光束整形进行原理仿真，并利用680 nm激光建立了实验系统进行光束整形验证。采用光束填充因子、光场调制度、均方根误差三种评价参数对整形结果进行评价，其中单平顶和多平顶整形的光束填充因子由整形前的36.1%分别提高到62.3%和56.7%，光场调制度由73.3%分别降到25.6%和30.3%。最后对采用数字微镜阵列进行光束整形的能量利用率进行了测量和分析。 |
| 英文摘要 | Two-photon microscopy(TPM) imaging technology combines confocal laser scanning technique and two-photon excitation technique. Due to deeper imaging, little photobleaching and phototoxicity, TPM is especially suitable for living imaging of cells, embryos and tissues, and has been widely used in life sciences. In this paper, a two-photon microscopy system has been set up; the group delay dispersion(GDD) was then measured and compensated; moreover, two-photon imaging and multi-focus rapid scanning have been done. Firstly, the inverted TPM imaging system was set up. The conjugated relation, object-image relation and the selection of system components were analyzed. The two-photon system control process and function of imaging software were also introduced. Secondly, to improve the two-photon excitation efficiency ,GDD was corrected. With the autocorrelator, the pulse width of femtosecond laser was measured, from four positions of optical path, and also optimal GDD compensation value of five wavelengths was measured, from the sample’position, and those data were fitted to obtain the full-band GDD compensation curve. Experimental results show the pulse width of sample position is decreased averagely 95fs after dispersion compensation, and that fluorescence intensity of biological sample is increased by 42.7% and 76.8% in two typical excitation wavelengths (700 nm and 900 nm) respectively. The linear inverse relationship between two-photon excitation efficiency and the pulse width of femtosecond laser is verified.Thirdly, flat-top beams spatial shaping of high power laser has been investigated. In this paper, we simulated the formation of single flat-top and multi-flattop beams by digital micromirror device and error diffusion method. An experimental setup with 680nm laser has been built to validate the model. Using beam fill factor, light field modulation, root mean square error as parameters to evaluate the beam shaping result, the beam fill factors increase from 36.1% to 62.3%(single)、56.7%(multi) respectively, while the light field modulation factors decrease from 73.3% to 25.6%(single)、30.3%(multi) respectively. Finally, energy efficiency of beam shaping using digital micromirror device was measured and analyzed. |
| 语种 | 中文 |
| 公开日期 | 2016-05-03 |
| 内容类型 | 学位论文 |
| 源URL | [http://ir.ciomp.ac.cn/handle/181722/49250]  |
| 专题 | 长春光学精密机械与物理研究所_中科院长春光机所知识产出 |
| 推荐引用方式 GB/T 7714 | 娄艳阳. 双光子显微成像系统关键技术研究[D]. 中国科学院大学. 2015. |
| 个性服务 |
| 查看访问统计 |
| 相关权益政策 |
| 暂无数据 |
| 收藏/分享 |
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。
修改评论