Development of an autostage cytofluorometer combined with an fluorescence image analyzer

结合荧光图像分析仪的自动台细胞荧光计的研制

• 批准号: 60870016
• 负责人: ASHIHARA Tsukasa
• 金额: $ 2.88万
• 依托单位: Kyoto Prefectural University of Medicine, Department of Pathology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60870016/
• 关键词: Cytofluorometry; Dual fluorescence measurement; Fluorescence image analysis; Autostage; DNA; Cancer cell diagnosis; 形態定量解析; 細胞測光; 螢光多重測光; 自動ステージ測光; 画像解析; 癌細胞診断法

Microscope-based cytofluorometry has been widely used for analyzing cell function (cell proliferation and differentiation) based on the measurement of each cellular macromolecule content (DNA, RNA, protein etc.). On the other hand, the image processing has recently been greatly progressed for both instruments as well as softwares, but presently this technique still seems difficult for the quantitative analysis of cell morphology since the cell functional significance cannot be easily understood by this technique alone. The proposed project was undertaken to develop a novel cell analysis system in which the combined analysis of cell function and cell morphology can be made using the fluorescence stained cells in smears. As a result, we developed an autostage cytofluorometer system (NIKON Pl-Auto-1) to which an image processor (ADS PIP-4000) with a monochromatic, high sensitivity TV camera was connected. The entire data processing was designed to carry out with a personal computer (NEC PC-9801 Vm2) through RS-232C and GP-IB interfaces. The softwares were constructed by combining our previously made programs for cytofluorometry (BASIC) and the packaged programs for the image processing (assembler), and the combined analysis of both the cellular fluorescence intensity and image was simultaneously achieved for the individual cells using an autostage scanning. It became also possible to quantitate the cell image intensity (i.e., to measure the amount of the stained materials) by integrating the pixel values for the selected wavelengths. Through the application experiments for human cancer cells, it was found that the combined analysis of DNA or DNA-RNA cytofluorometry and the "cell irregularity index" is very efficient for the diagnosis of malignancy. Studies are now under way to make our analysis method more practical.

基于显微镜的细胞荧光法已被广泛应用于分析细胞功能（细胞增殖和分化），该方法基于对每个细胞大分子（DNA、RNA、蛋白质等）含量的测量。另一方面，图像处理最近在仪器和软件方面都取得了很大的进展，但目前这项技术对于细胞形态的定量分析似乎仍然很困难，因为细胞的功能意义不能仅通过这项技术轻易地理解。本项目旨在开发一种新的细胞分析系统，利用荧光染色细胞的涂片进行细胞功能和细胞形态的综合分析。因此，我们开发了一种自动级细胞荧光计系统（尼康Pl-Auto-1），其中连接了带有单色高灵敏度电视摄像机的图像处理器（ADS PIP-4000）。整个数据处理被设计为通过RS-232C和GP-IB接口在个人计算机（NEC PC-9801 Vm2）上进行。该软件是通过结合我们之前制作的细胞荧光测定程序（BASIC）和打包的图像处理程序（汇编程序）构建的，并且使用自动级扫描同时实现对单个细胞的细胞荧光强度和图像的组合分析。还可以通过积分所选波长的像素值来量化细胞图像强度（即测量染色材料的数量）。通过对人癌细胞的应用实验，发现DNA或DNA- rna细胞荧光法与“细胞不规则指数”联合分析对恶性肿瘤的诊断是非常有效的。目前正在进行研究，以使我们的分析方法更加实用。

项目成果

Katsuyuki KUSUZAKI: "DNA-RNA cytofluorometric analysis of the cell proliferation of the bone giant cell tumors in relation to histopathology" Jap. J. Orthopaed, Surg. Assoc. (in Japanese). 60 (1). 51-60 (1986)

Katsuyuki KUSUZAKI：“与组织病理学相关的骨巨细胞瘤细胞增殖的 DNA-RNA 细胞荧光分析”

臨床検査. 29-2. (1985)

临床检查。29-2。

楠崎克之: 日整会誌. 60(1). 51-60 (1986)

楠崎克之：日本经济杂志 60(1) 51-60 (1986)。

Tsukasa ASHIHARA: "Development of an autostage cytofluorometer combined with fluorescence image analyzer" J. Histochem. Cytochem.

Tsukasa ASHIHARA：“开发自动阶段细胞荧光计与荧光图像分析仪相结合”J. Histochem。

Ashihara, T.: "Multiparametric analysis iusing autostage cytofluorometry" Acta Histochem. Cytochem.19 (1). 51-59 (1986)

Ashihara, T.：“使用自动阶段细胞荧光测定法进行多参数分析”Acta Histochem。