Developmental Research for Confocal Laser Microscope to Realize Fluorescence Microtomography.

组织病理学的真正信息存在于三维空间中，无论是在细胞水平上还是在组织结构中。然而，到目前为止，大多数关于微观形态的信息都局限在二维空间内。众所周知，癌症的诊断是基于腺体轮廓的结构非典型性、细胞核的组织或染色质分布等，它们都是三维的。到目前为止，为了满足准3D感觉，已经采用了倾斜对象的立体观看。然而，这种方法与荧光材料在3D空间中的定量信息无关。为了克服这些问题，使用共聚焦显微镜的荧光显微层析成像是最合适的。本研究的目的是建立一个共聚焦显微镜系统，以实现荧光显微透射图的半自动定量。我们是与奥林普…研究小组合作开发的S光学股份有限公司开发的一种共聚焦荧光显微镜系统，可以半自动获得半透明物体的连续光学切片。这一系列图像存储在激光光盘中，可以随时检索，以构建从任何角度观看的立体图。然而，每个断层摄影术代表了该平面中荧光材料的定量方面，因此像素值的总和产生了该平面或该结构中存在的全部材料的值。研究结果表明，本文所构建的共焦系统在未来的三维物体研究中是有用的和强大的。然而，与发展了100多年的普通光学显微镜相比，该系统的研究时间仍然很短，需要进一步的改进才能完善和实现定量三维显微镜。较少

True information in histopathology resides in tree-dimensional space, either in cellular level or in histological structures. Until present, however, most information on microscopic morphology is limited within two-dimensional space. Cancer diagnosis, has been, as well known, based on structural atypism of glandular contour, organization of cell nuclei, or chromatin distribution, etc. All of them are three-dimensional. So far stereoscopic viewing by tilting the object has been adopted to satisfy quasi-3D feeling. This method, however, is nothing to do with quantitative information of fluorescent materials in the 3D space. This stereoscopy is really superficial.To overcome these problems, flurorescence microtomography by using a confocal microscope is the most suitable. The present investigation aims at building a confocal microscope system to realize semi-automatically the fluorescence micro tromography in quantitative terms.We developed with colloboration with Research Group of Olympu … More s Optic Co. A confocal fluorescent microscope system which can obtain serial optic sections of semitransparent object, semi-automatically. The series of images are stored in laser disc and can be retrieved at any time to construct stereograms viewed at any angle. Each tomograph, however, represents quantitative aspect of fluorescent materials in that plane, so that summation of the pixel values produce the value of total material present in that plane or in that structure. This system is proved useful for application of quantitation of aimed materials distributed in 3D space in microscopic object.Results of the research, summarized above, indicates that the confocal system constructed in the present developmental research is useful and powerful in future investigation of 3D object. Nevertheless, further improvement would be necessary to perfect and accomplish quantitative 3D microscopy, since the time spent for investigation on this system is still short in comparison with usual light microscope which had been developed over 100 and more years. Less