| Research Abstract |
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
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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
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