Development of Nano-space Video Microscopy and its Application to the Analysis of New Pharmacological Dyanamics

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1) Development of New Video-microscopes for Nano-space ResolutionThe resolution of video microscope was improved by video enhanced contrast method and using insulators to get 60nm. Agold colloid in10nm scale was successfully observed by the dark field microscopy whose condenser lens was developed by Olympus Co.,2) Development of a Video Microscopy for Imaging of Fluorescence DepolarizationIn order to visualize the molecular mobility, one set of polarizers(parallel and perpendicular)was alternatively changed by rotating filter cassette in optical path of afluorescence video microscope. The molecular movement in the cell membrane was analyzed by image processing3) Development of Laser Scaling Video-microscopy : The interference fringe pattern at the crossing point of two crossed laser beams was found to be detected by video-microscopy. This fringe can be used as the optical scale for the evaluation of 2-&3-dimensional shapes of cells.Development of Analytical Methods for New Pharmacological Dynamics :4) Using high resolution video microscope, the role of cytoskeleton molecules in the sequence of phagocytosis and exocytosis of aneutrophil was measured. The effect of various medicinal molecules on the time course of phago-and exo-cytosis were analyzed to show the cytoskeleton molecules have akey role for it. Using fluorescence depolarization video-microscopy, it was shown that the membrane molecules of small intestinal epitheliums were squirming when the absorption promoting molecules have attacked on it. The combination of pulse flow injection method and microscopic observation, the beating of bronchial cilium in the presence and absence of expectorants was monitored.It was shown that the beating was promoted by Na-ambroxol.The developed microscopies are found to be new tools for the analysis of pharmacological dynamics of drugs.