基于光纤的混合照明光学层析荧光显微成像研究
发布时间：2019-02-11 13:35
【摘要】：荧光显微镜是生命科学研究中的重要工具。在普通的宽场荧光显微成像中,焦外的信号也会被探测器收集,严重降低成像对比度和分辨率。能够克服焦外干扰的显微镜(如共聚焦、双光子显微镜)往往成像速度较慢,设备昂贵。基于散斑和均匀光混合照明的光学层析方法成本低,层析效果好,在小型化的设备上很有应用前景。传统的散斑和均匀光混合照明光学层析方法使用自由空间中的散射片散射激光来生成散斑,光路固定且所占体积较大,不利于与商业显微镜集成。本文利用多模光纤导光及振动光纤消散斑方法,改进了传统的混合照明层析方法,实现了一种基于光纤的混合照明光学层析荧光显微成像系统。该系统的层析能力能够接近共聚焦显微成像;且具有光路封闭、装置空间柔性、系统简单、成本低(两万元左右),适合与商用显微镜结合等优点。本文主要工作如下:首先,给出了光纤导光的混合照明光学层析荧光显微成像系统原理图;对散斑和均匀光混合照明光学层析方法进行了理论分析,给出了具体算法;分析了振动光纤消散斑实现均匀照明的方法。其次,通过实验测试了不同参数对振动光纤消散斑效果的影响:振动电机驱动电压越高,消散斑效果越好,并且随着电压升高提升效果越弱;光纤模式数越大,消散斑效果越好;光纤缠绕不提升消散斑效果,甚至有微小的恶化。根据实验结果,设计了振动光纤消散斑模块及其工程封装,得到一个即插即用、调节方便、成本低、可用于各类光学系统的消散斑模块。最后,搭建了基于光纤的散斑和均匀光混合照明成像系统;编写了Matlab图像处理程序;利用花粉粒和100μm厚小鼠脑片样品对本方法进行了测试,得到了显著的层析效果。
[Abstract]:Fluorescence microscope is an important tool in life science research. In conventional wide-field fluorescence microscopic imaging, out-of-focal signals are also collected by detectors, which seriously reduce imaging contrast and resolution. Microscope, such as confocal microscope and two-photon microscope, which can overcome the external focal interference, is often slow and expensive. The optical tomography method based on speckle and uniform illumination has low cost and good performance. It has a good prospect in miniaturization equipment. The traditional optical tomography method of mixed illumination with speckle and uniform light uses scattering laser in free space to generate speckle. The light path is fixed and the volume is large, which is not conducive to the integration with commercial microscope. In this paper, the traditional hybrid illumination tomography method is improved by using multimode optical fiber light guide and vibratory optical fiber dissipation method, and a hybrid illumination optical tomography fluorescence microscopic imaging system based on optical fiber is implemented. The system has the advantages of close optical path, flexible device space, simple system, low cost (about 20,000 yuan), and suitable for combining with commercial microscope. The main work of this paper is as follows: firstly, the principle diagram of hybrid illumination optical tomography fluorescence microscopic imaging system is given, and the theoretical analysis and algorithm of speckle and uniform illumination optical tomography are given. The method of realizing uniform illumination of vibratory fiber dissipation spot is analyzed. Secondly, the effect of different parameters on the effect of vibration fiber dissipation is tested: the higher the driving voltage of vibration motor, the better the effect of dissipating spot, and the weaker the lifting effect is with the increase of voltage; The larger the number of fiber modes, the better the effect of dissipating speckle, and the less the effect of fiber winding is, the worse the effect is. Based on the experimental results, the vibratory fiber dissipation module and its engineering package are designed. A plug and play module with convenient adjustment and low cost is obtained, which can be used in various optical systems. Finally, a hybrid illumination imaging system based on fiber optic speckle and uniform light was set up, Matlab image processing program was compiled, and the method was tested by pollen grains and 100 渭 m thick mouse brain.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：TP391.41;TH742
本文编号：2419776
[Abstract]:Fluorescence microscope is an important tool in life science research. In conventional wide-field fluorescence microscopic imaging, out-of-focal signals are also collected by detectors, which seriously reduce imaging contrast and resolution. Microscope, such as confocal microscope and two-photon microscope, which can overcome the external focal interference, is often slow and expensive. The optical tomography method based on speckle and uniform illumination has low cost and good performance. It has a good prospect in miniaturization equipment. The traditional optical tomography method of mixed illumination with speckle and uniform light uses scattering laser in free space to generate speckle. The light path is fixed and the volume is large, which is not conducive to the integration with commercial microscope. In this paper, the traditional hybrid illumination tomography method is improved by using multimode optical fiber light guide and vibratory optical fiber dissipation method, and a hybrid illumination optical tomography fluorescence microscopic imaging system based on optical fiber is implemented. The system has the advantages of close optical path, flexible device space, simple system, low cost (about 20,000 yuan), and suitable for combining with commercial microscope. The main work of this paper is as follows: firstly, the principle diagram of hybrid illumination optical tomography fluorescence microscopic imaging system is given, and the theoretical analysis and algorithm of speckle and uniform illumination optical tomography are given. The method of realizing uniform illumination of vibratory fiber dissipation spot is analyzed. Secondly, the effect of different parameters on the effect of vibration fiber dissipation is tested: the higher the driving voltage of vibration motor, the better the effect of dissipating spot, and the weaker the lifting effect is with the increase of voltage; The larger the number of fiber modes, the better the effect of dissipating speckle, and the less the effect of fiber winding is, the worse the effect is. Based on the experimental results, the vibratory fiber dissipation module and its engineering package are designed. A plug and play module with convenient adjustment and low cost is obtained, which can be used in various optical systems. Finally, a hybrid illumination imaging system based on fiber optic speckle and uniform light was set up, Matlab image processing program was compiled, and the method was tested by pollen grains and 100 渭 m thick mouse brain.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：TP391.41;TH742
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