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视频探测器可以探测到显微镜样品台下方的两束激光交叉后形成的微区干涉条纹。在条纹图案照射下的生物样品图像可用于分析样品的尺度和三维形状，例如细胞。由几根光纤引入的多光束也被照射到一个细胞中。该方法具有对细胞辐照损伤小、同时获得不同波长荧光图像的优点。2)应用视频纳米显微镜分析大鼠粘膜肥大细胞RBL-2H3肿瘤类似物的细胞动力学和细胞对药物的反应。观察胞吐瞬间，分析胞吐数的时间过程。用钙离子载体A23187刺激细胞时发现两个峰，反映了细胞内的信号转导过程包含两个分离的过程。利用抑制剂或药物分子，可以更清晰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也被用于分析胰岛素分泌过程。对微颗粒的胞吐过程也进行了可视化，并对这一过程进行了定量分析。3)将该视频图像分析方法应用于其他领域的分离过程，并通过自制的宏观摄像机进行了成功的监控。对带移动和捕获过程的分析揭示了长期以来一直是黑盒子的柱式液相色谱分离过程的基本方面。

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.