Morpho-Physiological Study on Mechanism of Arrhythmia in Heart Muscle Cell by Using High-Speed Image Processor

利用高速图像处理器对心肌细胞心律失常机制进行形态生理学研究

• 批准号: 02670159
• 负责人: TAKAMATSU Tetsuro
• 金额: $ 1.41万
• 依托单位: Kyoto Prefectural University of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02670159/
• 关键词: Arrhythmia; Heart Muscle Cell; Calcium Wave; Confocal laser Scanning Microscope; Image Processing; Indoー1

I have developed a dual, digital fluorescence imaging system and new confocal laser scanning microscope employing an inverted optical system for imaging fast dynamic changes of the intracellular calcium ion concentration ([Ca^<2+>]_i) in heart muscle cells.In 1990, Images of[Ca^<2+>]_i taken at 67-ms intervals during propagation of calcium waves revealed that the calcium wave front was constant in shape, spatially steep, typically rising from 500 to 1200 nM in about 10 mu m, and propagating at constant velocity, typically 100 mu m/sec at 22 ﾟC. In 1991, I have investigate the biological application of a confocal laser scanning microscope for imaging the calcium wave. This instrument has several advantages for observing moving objects and flexible choice of scanning modes. 1) A faster scanning speed of up to 0.25 sec per frame including line scan, fast horizontal and vertical full-frame scan and a faster horizontal narrow band scan. 2) An efficient optical path specially designed for th … More e inverted fluorescence microscope system. 3) Direct transfer of digitized images from the frame memory in the CLSM to that of an image processor, avoiding the degradation of densitometric data by duplicated A/D and D/A conversion. These features gave us the opportunity to investigate the properties of the calcium wave during its propagation between isolated guinea-pig ventricular cell pairs. The actual[Ca^<2+>]_i is obtained directly through the fluorescence intensity of injected fluo-3, which responds to changes of[Ca^<2+>]_i in optically sectioned unit volumes of the cell. Optical sectioning images obtained enable us to determine the cell-to-cell junction between heart muscle paired cells and reveal that the high[Ca^<2+>]_i can evoke calcium-induced calcium release from the sarcoplasmic reticulum in the neighboring heart muscle cells through the gap junction without delay.The confocal laser scanning microscope with depth-discriminating ability is a valuable tool for taking pictures of the sequence of biological events in living heart muscle cells such as the calcium wave, a model for arrhythmia at cellular level. Less

我开发了一种双数字荧光成像系统和新型共焦激光扫描显微镜，采用倒置光学系统对心肌细胞内钙离子浓度 ([Ca^<2+>]_i) 的快速动态变化进行成像。 1990 年，在钙波传播期间以 67 毫秒间隔拍摄的 [Ca^<2+>]_i 图像显示，钙波前是 形状恒定，空间陡峭，通常在约 10 μm 内从 500 nM 上升到 1200 nM，并且以恒定速度传播，通常在 22 °C 时为 100 μm/sec。 1991年，我研究了共焦激光扫描显微镜在钙波成像方面的生物学应用。该仪器对于观察运动物体具有多种优势，并且可以灵活选择扫描模式。 1) 更快的扫描速度，每帧高达 0.25 秒，包括行扫描、快速水平和垂直全帧扫描以及更快的水平窄带扫描。 2) 专为倒置荧光显微镜系统设计的高效光路。 3) 将数字化图像从 CLSM 中的帧存储器直接传输到图像处理器的帧存储器，避免由于重复的 A/D 和 D/A 转换而导致密度数据的退化。这些特征使我们有机会研究钙波在离体豚鼠心室细胞对之间传播期间的特性。实际的[Ca^<2+>]_i直接通过注入的fluo-3的荧光强度获得，其响应细胞光学切片单位体积中[Ca^<2+>]_i的变化。获得的光学切片图像使我们能够确定心肌配对细胞之间的细胞与细胞连接，并揭示高[Ca^<2+>]_i可以通过间隙连接立即引起钙诱导的钙从邻近心肌细胞的肌浆网释放。具有深度辨别能力的共焦激光扫描显微镜是拍摄心肌细胞序列的有价值的工具。 活心肌细胞中的生物事件，例如钙波，细胞水平的心律失常模型。较少的

项目成果

南川 哲寛,高松 哲郎,河内 秀幸,藤田 晢也: "共焦点レ-ザ走査顕微鏡を用いた単離心筋細胞のカルシウム波の解析法" 医学のあゆみ. 155. 473-474 (1990)

Tetsuhiro Minamikawa、Tetsuro Takamatsu、Hideyuki Kawachi、Akiya Fujita：“使用共焦激光扫描显微镜分析分离心肌细胞中钙波的方法”医学史 155. 473-474 (1990)。

高松 哲郎,南川 哲寛,藤田 晢也: "組織細胞化学 レ-ザ顕微鏡の原理,技術と医学生物学への応用" 学際企画, 196 (1991)

Tetsuro Takamatsu、Tetsuhiro Minamikawa、Akiya Fujita：“组织细胞化学：激光显微镜原理和技术及其在医学生物学中的应用”跨学科规划，196（1991）

藤田 晢也,高松 哲郎,南川 哲寛: "限界を超える生物顕微鏡(宝谷 紘一・木下 一彦編) 4章 立体断面を見る(共焦点レ-ザ-走査蛍光顕微鏡)" 学会出版センタ-, 177 (1991)

Akiya Fujita、Tetsuro Takamatsu、Tetsuhiro Minamikawa：“生物显微镜超越极限（Koichi Takaraya 和 Kazuhiko Kinoshita 编辑）第 4 章：查看 3D 横截面（共焦激光扫描荧光显微镜）”Gakkai 出版中心，177（1991））

Tetsuro Takamatsu,Tetsuhiro Minamikawa,Hideyuki Kawachi and Setsuya Fujita: "Imaging of Calcium Wave Propagation in GuineaーPig Ventricular Cell Pairs by Confocal Laser Scanning Microscopy" CELL STRUCTURE AND FUNCTION. 16. 341-346 (1991)

Tetsuro Takamatsu、Tetsuhiro Minamikawa、Hideyuki Kawachi 和 Setsuya Fujita：“通过共聚焦激光扫描显微镜对豚鼠心室细胞对中的钙波传播进行成像”《细胞结构和功能》16. 341-346 (1991)。

Tetsuhiro Minamikawa,Tetsuro Takamatsu and Setsuya Fujita: "Application of Laser Microtomography to in Situ ThreeーDimensional Subcellular Morphology" ACTA HISTOCHEM.CYTOCHEM.24. 55-60 (1991)

Tetsuhiro Minamikawa、Tetsuro Takamatsu 和 Setsuya Fujita：“激光显微断层扫描在原位三维亚细胞形态学中的应用”ACTA HISTOCHEM.CYTOCHEM.24（1991）。