双光子显微成像实验及理论分析研究
本论文主要分为四章。第一章从显微镜的发展历史入手，介绍了一些常见的显微镜，并比较了几种显微镜的区别。第二章立足于双光子的作用机理，从数值上具体确立了双光子显微镜的优点。第三章介绍双光子显微镜的具体构成部分：光学成像系统、扫描系统、光电转换系统。主要讨论了利用光自准直方法，对激光光束偏移的测量和修正的光路。第四章利用仪器分别对植物细胞单光子成像和双光子成像，直观上体现双光子成像的优势。26423
关键词 双光子 荧光 显微镜 共聚焦 自准直
毕业论文设计说明书外文摘要
Title Experimental and Theoretically Analysis of Two-photon Microscopy
Abstract
The mainly working principal of two-photon microscopy is the two-photon fluorescence effect. The infrared light or near infrared light transmitted by femtosecond laser is incident on the fluorophores. The simultaneously absorption of two incident photons leads to the two-photon excitation of fluorescence, which used for the 3D(three dimension) imaging.This kind of microscope brings many advantages, such as reduction of the light damage, enhanced resolution and imaging depth.
This thesis is constructed by four parts. Chapter one has a brief introduction of the history of microscopes, introduces some regular microscopes and makes a compare between them. Chapter two explains the two-photon fluorescence effect theoretically and proves the advantages numerically. Chapter three introduces the mainly components: imaging system, scanning system and photovoltaic conversion system. The optical path detecting and compensating the light beam drift, which relied on the autocollimation technology are mainly discussed. Chapter four contains the the imaging process of one-photon and two-photon fluorescence,at the same time the advantages of two-photon fluorescence are directly displayed.
Keywords two-photon fluorescence microscope confocal autocollimation
目 次
1 绪论 1
1.1 各类显微镜的介绍3
1.2 几种显微镜的比较5
1.3 全文结构布局安排6
2 双光子荧光显微成像7
2.1 双光子显微镜的简介 7
2.2 双光子激发基本理论 8
2.3 双光子显微成像优缺点 9
2.4 双光子显微成像中的主要参数 9
3 双光子显微镜系统简介 15
3.1 实验装置简介15
3.2 光学成像系统 15
3.3 激光扫描系统20
3.4 光电转换系统22
4 实验 25
4.1 实验仪器 25
4.2 实验过程 26
4.3 实验参数 26
4.4 实验结果 27
结论 28
致谢 29
参考文献30
1 绪论
人眼是人类观察世界的主要窗口，通过这个窗口人们可以看到这个五彩缤纷的世界。然而，人类对世界的认知只能限制在肉眼的程度，由于眼睛有限的分辨率（<0.1mm的物体无法观察），所以对于微观世界的观察是不可能。随着时代的变迁，于是出现了最简单的辅助观察的手持透镜，即所谓的“放大镜”，它利用光线偏折形成放大的虚像。然而单个凸透镜的放大倍率只有几十倍，仍然没有办法观察微米级的物体细节，于是显微镜变应运而生。16世纪，荷兰的Janssen发明了第一台显微镜，它由几个镜片组成（物镜，目镜），放大率达到了200，从而使得人类进入了微观世界。