激光共焦扫描荧光显微内窥镜研究
发布时间：2019-04-20 13:18
【摘要】：激光共焦扫描显微内窥镜凭借其能实现高分辨率的实时组织学诊断和一定深度的断层扫描成像,已经成为当前研究的热点。医生通过激光共焦扫描显微内窥镜不仅能观察人体内脏器官表面的组织形态,而且可以深入表皮细胞无需取样即可诊断病变情况,还可以使用荧光对比剂,特异性强,操作简单。目前,激光共焦扫描内窥镜虽然已经有少数商用产品面市,但是多采用光纤束方案,鲜有国内产品,因此研究激光共焦内窥镜有重要意义 本文研制了一种激光共焦扫描荧光显微内窥镜,采用望远式显微内窥光学系统,同时实现长距离的图像中继传输、远心f-theta光学扫描和显微内窥成像功能。X,Y方向共焦扫描由双振镜实现,Z轴扫描由压电马达实现,低噪声扫描控制信号由嵌入式系统产生,通过三者结合完成三维共聚焦扫描。激光共焦扫描显微内窥镜采用小型化设计方案,主要工作内容包括以下三部分：(一)设计完成大视场、大数值孔径、长距离图像中继传输的显微内窥成像光学系统,将激光耦合进体内组织,并将荧光信号耦合至体外探测器,在获得高分辨率和高对比度荧光图像的同时,缩小内窥镜探头直径。最终探头外径尺寸为8 mm,工作长度为250.3 mm,可通过标准腹腔镜手术孔进行体内显微内窥成像；(二)采用3 mm通光孔径的小尺寸平面反射镜实现体外共焦扫描,摆动频率为100 Hz,扫描范围达到600μm,由嵌入式系统控制实现X,Y快速共焦扫描；(三)样品含有荧光试剂,激光激发后高灵敏度PMT器件探测并成像,上传至PC实现图像重建和处理。激光控制和荧光探测仅通过电缆和光纤与共焦扫描显微内窥镜前端连接,将扫描机构放置在体外,减小了显微内窥镜的前端尺寸和重量。 通过实验验证,本系统的成像视场为(?)600μm,光学分辨率为2.2μm,获得图像分辨率为512*320,每4S采集一帧图像。本系统可进一步发展为三维共焦显微成像系统,采用手持式或者其他方式工作,进行体内组织的共焦扫描成像,真正实现微创、在体的荧光显微内窥术。
[Abstract]:Laser confocal scanning microendoscopy (LSCM) has become the focus of current research because it can realize high-resolution real-time histology diagnosis and a certain depth of tomography imaging. By means of laser confocal scanning microendoscopy, doctors can not only observe the morphology of tissues on the surface of human internal organs, but also diagnose the lesions without sampling in the epidermis cells, and can also use fluorescent contrast media with strong specificity. The operation is simple. At present, although a small number of commercial products have been marketed for laser confocal scanning endoscopes, optical fiber bundle schemes are mostly used, and few domestic products are available. Therefore, it is of great significance to study laser confocal endoscopy. In this paper, a laser confocal scanning fluorescence microendoscope is developed, which uses telescopic endoscope optical system to realize long-distance image relay transmission. Telecentric f-theta optical scanning and microscopic endoscope imaging function. X, Y confocal scanning is realized by double vibroscope, Z axis scanning is realized by piezoelectric motor, low noise scanning control signal is generated by embedded system, and X, Y confocal scanning is realized by double vibroscope, Z axis scanning is realized by piezoelectric motor. The three-dimensional confocal scanning is accomplished by the combination of three-dimensional scanning. The laser confocal scanning microendoscope adopts the miniaturization design scheme. The main contents of the work include the following three parts: (1) the micro-endoscope optical system with large field of view, large numerical aperture and long-distance image relay transmission is designed and implemented. The laser is coupled into the tissue of the body and the fluorescence signal is coupled to the external detector to obtain high-resolution and high-contrast fluorescence images while reducing the diameter of the endoscope probe. The final external diameter of the probe is 8 mm, the working length is 250.3 mm, and the in vivo microscopic endoscopy can be performed by standard laparoscopic surgery. (2) the external confocal scanning is realized by using a small size planar reflector with 3 mm aperture. The swing frequency is up to 600 渭 m in the scanning range of 100 Hz,. The X and Y fast confocal scanning are realized by the embedded system control. (3) the sample contains fluorescence reagent. After laser excitation, the high sensitivity PMT device is detected and imaged, and the image is reconstructed and processed by uploading to PC. Laser control and fluorescence detection are connected to the front end of confocal scanning microendoscope only by cable and fiber, and the scanning mechanism is placed in the outer body, thus reducing the size and weight of the front end of the microendoscope. The experimental results show that the field of view of the system is (?) 600 渭 m, the optical resolution is 2.2 渭 m, the resolution of the image is 512 渭 m, and one frame of image is collected every 4S. This system can be further developed into a three-dimensional confocal microscopic imaging system, which can perform confocal scanning imaging of the tissues in vivo by hand-held or other methods, and truly realize micro-invasive, in-vivo fluorescence microscopic endoscopy.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：TH776.1;TN249
本文编号：2461640
[Abstract]:Laser confocal scanning microendoscopy (LSCM) has become the focus of current research because it can realize high-resolution real-time histology diagnosis and a certain depth of tomography imaging. By means of laser confocal scanning microendoscopy, doctors can not only observe the morphology of tissues on the surface of human internal organs, but also diagnose the lesions without sampling in the epidermis cells, and can also use fluorescent contrast media with strong specificity. The operation is simple. At present, although a small number of commercial products have been marketed for laser confocal scanning endoscopes, optical fiber bundle schemes are mostly used, and few domestic products are available. Therefore, it is of great significance to study laser confocal endoscopy. In this paper, a laser confocal scanning fluorescence microendoscope is developed, which uses telescopic endoscope optical system to realize long-distance image relay transmission. Telecentric f-theta optical scanning and microscopic endoscope imaging function. X, Y confocal scanning is realized by double vibroscope, Z axis scanning is realized by piezoelectric motor, low noise scanning control signal is generated by embedded system, and X, Y confocal scanning is realized by double vibroscope, Z axis scanning is realized by piezoelectric motor. The three-dimensional confocal scanning is accomplished by the combination of three-dimensional scanning. The laser confocal scanning microendoscope adopts the miniaturization design scheme. The main contents of the work include the following three parts: (1) the micro-endoscope optical system with large field of view, large numerical aperture and long-distance image relay transmission is designed and implemented. The laser is coupled into the tissue of the body and the fluorescence signal is coupled to the external detector to obtain high-resolution and high-contrast fluorescence images while reducing the diameter of the endoscope probe. The final external diameter of the probe is 8 mm, the working length is 250.3 mm, and the in vivo microscopic endoscopy can be performed by standard laparoscopic surgery. (2) the external confocal scanning is realized by using a small size planar reflector with 3 mm aperture. The swing frequency is up to 600 渭 m in the scanning range of 100 Hz,. The X and Y fast confocal scanning are realized by the embedded system control. (3) the sample contains fluorescence reagent. After laser excitation, the high sensitivity PMT device is detected and imaged, and the image is reconstructed and processed by uploading to PC. Laser control and fluorescence detection are connected to the front end of confocal scanning microendoscope only by cable and fiber, and the scanning mechanism is placed in the outer body, thus reducing the size and weight of the front end of the microendoscope. The experimental results show that the field of view of the system is (?) 600 渭 m, the optical resolution is 2.2 渭 m, the resolution of the image is 512 渭 m, and one frame of image is collected every 4S. This system can be further developed into a three-dimensional confocal microscopic imaging system, which can perform confocal scanning imaging of the tissues in vivo by hand-held or other methods, and truly realize micro-invasive, in-vivo fluorescence microscopic endoscopy.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：TH776.1;TN249
【引证文献】
相关博士学位论文 前1条
1 雷翔;板条固体激光器光束净化控制技术研究[D];中国科学院研究生院（光电技术研究所）;2013年
，本文编号：2461640
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