Studies for objective diagnosis of malignancy in biopsy materials by nuclear DNA quantification and image analysis

人类病理学家必须在他们检查的每一个活组织检查中建立良性或恶性性质的诊断，并感到有必要引入一些客观的线索来确定最终的决定。这种帮助的可能候选是细胞的DNA测量和其形态分析。1985年，将硅增强管（SIT）摄像机（Hamamatsu Photonics）与落射荧光显微镜（Vannox，Olympus）连接，首先测定DAPI染色的细胞核的荧光，然后用SIT摄像机拍摄相同细胞的图像。对细胞图像的每个像素进行积分，并将数据与细胞荧光测量的数据进行比较。重复这个过程，我们可以建立相机平衡，电压范围，滤波器选择的最佳条件，并开发软件来最有效和正确地驱动这个系统。1986年，我们将该系统应用于胃、结肠肿瘤细胞恶性程度的分析，发现用该系统定量的DNA含量的非整倍体和非典型形态，可对可疑活检中倾向于早期转移或浸润的癌细胞作出客观判断。结合凝集素染色和其他可识别或定量的癌细胞标志物，对建立正确的癌症预后有很大帮助。

Human pathologists must establish diagnosis of benign or malignant nature in every biopsy they examine,and feel the necessity of introducing some objective cues in determination of the final decision. Possible candidate of this help is DNA measurement of cells and analysis of their morphology. This research is aimed at developing a system to measure DNA content on fluorescent images of the cells taken by a silicon intensifer tube(SIT) video camera.In 1985, a SIT camera(Hamamatsu Photonics) is connected to an epi-illumination fluorescent microscope (Vannox, Olympus) and, at first, determination of fluorescence on DAPI-stained cell nuclei, and then, images of the same cells are taken by the SIT camera. Every pixel of the cell image is integrated and the data is compared with that of the cytofluorometry. Repeating this procedure, we can establish optimal conditions of camera balancing, voltage range,selection of filters, and develop software to drive this system most efficiently and correctly. The program is added to the system in a form of ROM.In 1986, we apply this system to analysis of malignancy of tumor cells in the stomach and colon, and find that aneuploidy of DNA content and atypical shape quantified by this system give objective judgement of cancer cells that tend to metastasize or infiltrate early among suspicious biopsies. Combination of lectin staining and other cancer cell markers that can be identifed or quantified are of great help to establish correct prognosis of the cancer cases.