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目的制备猫视神经的连续切片,选择合适的染色方法 ,寻找合适的配准方法 ,经图像采集及初步处理后利用计算机的三维重建技术,获得可视化的猫视神经三维结构。方法利用石蜡切片技术的制备猫视神经连续切片,采用特殊的神经三色染色法进行切片染色。利用数码显微镜及Motic Images Advanced 3.0和Motic Images Assembly 1.0软件对连续的猫视神经石蜡切片进行拍照,获得猫视神经连续切片的原始图像,将原始图像在GIMP软件中经过调整亮度对比度、初步图像配准、提取神经束膜图像信息后在Amira软件下,经过计算机在手工配准的基础上进一步自动配准,把二维的切片图像重建成三维的可视化图像。结果实验所得经三色染色的连续切片的原始图像上神经束膜能较好的与周围的组织区分;利用Amira三维重建软件,重建得到猫视神经三维结构图像,重建后的猫视神经结构三维图像可进行任意的切割、旋转、回复、拍照等操作。结论实验证明在不利用标志线的条件下,通过制备视神经连续切片,经过染色拍照,手工图像配准后利用计算机自动配准技术对视神经进行三维重建是完全可行的。
OBJECTIVE To prepare continuous sections of cat optic nerve, select suitable staining methods and search for suitable registration methods. After image acquisition and initial processing, computerized three-dimensional reconstruction technique was used to obtain the visualized three-dimensional structure of cat optic nerve. Methods Serial slices of cat optic nerve were prepared by paraffin section technique and stained with special trichrome method. Continuous cat paraffin sections were photographed with digital microscope and Motic Images Advanced 3.0 software and Motic Images Assembly 1.0 software to obtain original images of continuous cat optic nerve slices. The original images were adjusted in GIMP software with brightness contrast, preliminary image registration, After extraction of the information of the perineurial image, it is further automatically registered by Amira software based on manual registration, and the two-dimensional slice image is reconstructed into a three-dimensional visualized image. Results The neurofilament on the original images obtained by the trichrome-stained serial images can be distinguished from the surrounding tissues better. Using the Amira reconstruction software, the three-dimensional images of the cat optic nerve can be reconstructed. The three-dimensional images of the reconstructed cat’s optic nerve can be reconstructed Do any cutting, rotation, reply, take pictures and other operations. Conclusion Experiments show that it is feasible to reconstruct the optic nerve by using computer automatic registration technology after preparing serial sections of optic nerve, dyeing pictures and manual image registration without using the sign line.