Development of Video Nanoscopes Using Crossed Laser Beams for Tracing of Drug Molecules

1)利用交叉激光束研制视频纳米显微镜在显微镜样品台下方入射的两束激光交叉形成的微区干涉条纹图可被视频探测器探测到。生物样品在条纹图照射下的图像可用于分析样品的尺度和三维形状，G.细胞将多束光通过多根光纤引入到单个细胞上，也照射到细胞上。该方法的优点在于对细胞的辐射损伤较小，并且可以在不同波长下获得荧光图像。2）视频纳米显微镜在细胞动力学和细胞对药物分子的反应分析中的应用使用大鼠粘膜肥大细胞的肿瘤类似物RBL-2 H3。观察胞吐的瞬间，并分析胞吐次数的时间过程。用钙离子载体A23187刺激细胞后，细胞内出现两个峰，反映了细胞内信号转导过程中存在两个分离的过程。利用抑制剂或药物分子，这种方法可以更清楚地揭示信号转导过程。小鼠胰岛素瘤细胞的肿瘤类似物MIN-6也用于胰岛素分泌过程的分析。3）将这种视频图像分析方法应用于其它场分离过程，成功地实现了宏观视频监控。对谱带移动和捕获过程的分析揭示了长期以来一直被视为黑箱的柱液相色谱分离过程的本质。

1) Development of Video Nanoscopes Using Crossed Laser BeamsInterference fringe pattern in micro region which was formed by crossed two laser beams coming from below the sample stage of a microscope was found to be detected by a video detector. The image of biological samples illuminated by the fringe pattern can be used for the analysis of scale and 3-d shape of the samples, e. g. cells. Multi beams which was introduced by several optical fibers onto a single cell was also irradiated to a cell. The method is advantageous in giving less irradiation damage to cells and getting fluorescence images at various wavelengths simultaneously.2) Application of Video Nanoscopes to the Analysis of Cell Dynamics and Cell Response to Drug MoleculesA tumor analog of rat mucosal mast cells, RBL-2H3, was used. The momentof exocytosis was visualized and the time course of the number of exocytosis was analyzed. Two peaks were found when the dells was stimulated with Ca-ionophore A23187 which reflects the two separated processes are included in the signal transduction process in this cell. Using inhibitors or drug molecules, clearer signal transduction process are now under revealing with this method. A tumor analog of mouse insulinoma cells, MIN-6, was also used for the analysis of insulin secretion process. The exocytosis of micro granules was also visualized and quantitative analysis of this process is under profress.3) Application of this Video lmage Analyzing Method to Other FieldSeparating process was successful monitored by our developed macro-videoscope. Analysis of band movement and trapping process revealed the fundamental aspect of the separation process in column liquid chromatography which has been black boxes for long time.