基于光纤束的共聚焦荧光内窥成像研究

发布时间：2018-04-20 13:05

本文选题：内窥 + 荧光成像　；参考：《华中科技大学》2013年博士论文

【摘要】：由于人口增长和老龄化的加剧，以及抽烟，酗酒，缺乏锻炼，摄入高油脂食物等不良生活习惯，癌症近年来发生的比重越来越多。在发达国家癌症已经成为第一致死因素，在发展中国家癌症成为第二致死因素。研究表明，癌症多发生在上皮细胞的浅层，对癌症的诊断发现越早，成活率越高。所以对癌症的早期诊断已经越来越重要，对癌症早期诊断的各种设备也成为研究的热点。在这些成像诊断方法中，共聚焦荧光内窥系统可以在体实时提供亚细胞分辨率的结构信息，减少或者有可能取代组织活检，受到广泛的关注。本文就基于光纤束的共聚焦荧光内窥成像系统展开研究，并提出了基于傅里叶变换的多光谱荧光内窥成像系统。本文研制开发了一种基于光纤束的共聚焦荧光内窥成像系统。并在光学结构，所用主要元件，控制程序和后期数据处理部分都进行了详细的讨论。在共聚焦荧光内窥成像系统中使用CRS共振型振镜实现扫描，，取得4帧/秒1024×1024图像。在系统的控制电路中使用NI公司的商用采集卡和图形化编程软件Labview，使得我们能够关注系统的设计而不是繁琐的编程。首先使用标准分辨率板对其横向分辨率进行了测试得到其分辨率4.38微米。使用光栅样品测试系统成像区域为790微米。然后使用标准荧光小球和花粉颗粒检验了其对标准荧光样品的成像能力。最后对正常昆明小鼠的离体和在体组织进行了染色成像实验，验证了系统亚细胞分辨率对组织样品结构信息的分辨能力。在多光谱荧光内窥成像系统中，我们结合了光纤束内窥成像的灵活性和傅里叶变换光谱仪的各种优势，可以在体对各种生物器官和组织进行高光谱分辨率的成像。在系统中我们设计使用了迈克尔逊干涉仪结构，选用精度较高的纳米位移台对运动平面镜进行移动。在系统中取出一部分照明光导入迈克尔逊干涉仪中实现对运动平面镜位置的实时监测。该种结构的设计可以保证系统的精确性和长期的稳定性。使用现有的平移台，理论上可以提供的最高光谱分辨率约是4.02cm~(-1)。本文使用光谱稳定的氦氖激光器对其光谱分辨率进行了测量，和理论值有着较好的吻合。然后使用该系统对多色小球进行光谱成像验证了系统分辨多种荧光染料的能力。最后通过对深脑部位神经元进行光谱成像，展示了使用光纤束带来的内窥光谱成像能力。另外本文结合光纤束内窥功能和傅里叶变换光谱仪系统对标准样品和组织的激发光谱进行了测量，展示了使用激发光谱区分不同荧光基团的功能。在共聚焦荧光内窥成像系统中使用光纤束可以带来成像的灵活性，但是光纤束本身也有蜂窝状的固有噪声。多种图像处理方法被提出用于消除光纤束的像素化。本文对视觉效果较好的自然邻点插值重建方法进行了论述。
[Abstract]:As a result of population growth and aging, as well as smoking, alcoholism, lack of exercise, intake of fatty foods and other bad habits, cancer in recent years the proportion of more and more. Cancer has become the first cause of death in developed countries and the second leading cause of death in developing countries. Studies have shown that cancer occurs mostly in the superficial layer of epithelial cells, and the earlier the cancer is diagnosed, the higher the survival rate is. Therefore, the early diagnosis of cancer has become more and more important. Among these imaging diagnostic methods, confocal fluorescence endoscopy system can provide subcellular resolution information in vivo in real time, reduce or possibly replace tissue biopsy, so it has attracted wide attention. In this paper, the confocal fluorescence endoscope imaging system based on optical fiber beam is studied, and a multispectral fluorescence endoscope imaging system based on Fourier transform is proposed. In this paper, a confocal fluorescence endoscope imaging system based on optical fiber beam is developed. The optical structure, main components, control program and later data processing are discussed in detail. In the confocal fluorescence endoscope imaging system, the CRS resonator is used to scan, and 4 frames / s 1024 脳 1024 images are obtained. In the control circuit of the system, the commercial acquisition card of NI company and the graphical programming software Labvieware used, which enables us to pay attention to the design of the system rather than the tedious programming. First, the horizontal resolution is measured with standard resolution board and the resolution is 4.38 渭 m. The imaging area of the system using the grating sample test system is 790 渭 m. Then the imaging ability of the standard fluorescent sample was tested with standard fluorescent pellets and pollen particles. Finally, in vitro and in vivo tissues of normal Kunming mice were stained to verify the resolution of the subcellular resolution of the system to the structural information of tissue samples. In the multispectral fluorescence endoscope imaging system, we combine the flexibility of optical fiber beam endoscope imaging and the advantages of Fourier transform spectrometer, we can carry out high spectral resolution imaging of various biological organs and tissues in vivo. The Michelson interferometer structure is used in the system, and the high precision nanometer displacement table is used to move the moving plane mirror. In the system, a part of the illumination light is introduced into the Michelson interferometer to realize the real-time monitoring of the position of the moving plane mirror. The design of this kind of structure can guarantee the accuracy and long-term stability of the system. Using the existing translation table, the theoretical maximum spectral resolution is about 4.02 cm ~ (-1) cm ~ (-1) ~ (-1). In this paper, the spectral resolution of He-Ne laser with stable spectrum is measured, which is in good agreement with the theoretical value. Then the system was used for spectral imaging of polychromatic pellets to verify the ability of the system to distinguish various fluorescent dyes. Finally, spectral imaging of deep brain neurons is performed to demonstrate the ability of spectral imaging with optical fiber beams. In addition, the excitation spectra of standard samples and tissues were measured with the help of optical fiber beam endoscope and Fourier transform spectrometer system, and the function of using excitation spectra to distinguish different fluorescence groups was demonstrated. In confocal fluorescence endoscope imaging system, the use of optical fiber beam can bring the flexibility of imaging, but the fiber bundle itself also has honeycomb inherent noise. A variety of image processing methods are proposed to eliminate the pixel of optical fiber bundles. In this paper, the method of natural neighbor interpolation reconstruction with better visual effect is discussed.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R310

【共引文献】