Trial manufacture of an ultra-high sensitive video-microscope system with and image-processor and its applications to cell biology

开发了一种视频显微镜系统，以便在极低强度的光下观察微观物体。利用该系统，记录了海胆、海星、青鳉、青蛙和金黄地鼠卵受精和人工激活时细胞内钙离子的释放，这些卵被注射了对游离钙离子敏感的发光蛋白水母发光蛋白。在这些卵子中，细胞内钙离子浓度的增加开始于精子进入的部位或人工激活的部位，并在细胞皮层上向反足传播。这种钙波先于皮层肺泡或皮层颗粒的破裂波，表明钙离子的增加引起了皮层的变化。在注射水母发光蛋白的青鳉和海胆受精卵中记录到细胞内钙离子伴随细胞周期的周期性变化。在两种卵中，细胞内钙离子浓度在卵裂过程中均达到最低值，用水母发光蛋白（aequorin）作为探针，记录到细胞内钙离子浓度随黏菌的变形虫运动和原生质流而短暂和局部增加，用钙离子荧光探针fura 2也记录到细胞内钙离子浓度的变化。

A video-microscope system was developed, in order to visualize microscopic objects under an extremely-low intensity light. Using this system, intracellular release of calcium ions at fertilization and artifical activation was recorded in eggs of sea urchin, starfish, medaka, frog and golden hamster, which had been injected with aequorin, a photoprotein sensitive to free calcium ions. In these eggs, the increase in intracellular calcium ion concentration was started at the site of sperm entry or the site of artificial activation and it propagated over the cell cortex toward the antipod. This calcium wave preceded the wave of the breakdown of cortical alveoli or cortical granules, suggesting that the increase in calcium ions induces the cortical change.Periodic changes in intracellular calcium ions accompanying cell cycle were recorded in fertilized medaka and sea urchin eggs injected with aequorin. In both eggs, intracellular calcium ion concentration attained a minimum during cleavage.Transient and localized increases in intracellular calcium ions accoumpanying amoevoid movement and protoplasmic streaming in slime mold were recorded using aequorin as a probe.Changes in intracellular calcium concentration were also recorded using fura 2, a fluorescent probe for calcium ions.