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

纳米空间视频显微镜的发展及其在新药理动力学分析中的应用

• 批准号: 05557101
• 负责人: MASUJIMA Tsutomu
• 金额: $ 7.1万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05557101/
• 关键词: Microscopy; Video Analysis; Fluorescence Depolarization; Image Analysis; Screening for Drug; Pharmacological Dynamics; Cell Dynamics; ビデオスペクトロメーター

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.

1）开发纳米空间分辨率的新型视频显微镜通过视频增强对比度方法和使用绝缘体来提高视频显微镜的分辨率，达到60nm。利用奥林巴斯公司开发的聚光镜暗视野显微镜成功地观察到了10nm尺度的金胶体。2）荧光去偏成像视频显微镜的研制为了可视化分子迁移率，通过旋转荧光视频显微镜光路中的滤光片盒交替改变一组偏振器（平行和垂直）。通过图像处理分析细胞膜中的分子运动3) 激光缩放视频显微镜的发展：发现通过视频显微镜可以检测到两束交叉激光束交叉点处的干涉条纹图案。该条纹可用作评估细胞2维和3维形状的光学标尺。新药理动力学分析方法的开发：4）使用高分辨率视频显微镜，测量细胞骨架分子在嗜中性粒细胞吞噬和胞吐作用序列中的作用。分析了各种药物分子对吞噬和胞吐作用时程的影响，表明细胞骨架分子对其具有关键作用。使用荧光去偏视频显微镜显示，当吸收促进分子攻击小肠上皮细胞的膜分子时，其膜分子会蠕动。脉冲流注射法与显微镜观察相结合，对有或无祛痰剂时支气管纤毛的跳动进行了监测，发现氨溴索促进了支气管纤毛的跳动。所开发的显微镜为药物药理动力学分析提供了新的工具。

项目成果

小澤 孝一郎: "ビデオマイクロスコープを用いた細胞動態リアルタイム解析" 神経化学. 33. 204-205 (1994)

Koichiro Ozawa：“使用视频显微镜实时分析细胞动力学”《神经化学》33. 204-205 (1994)。

K.Ozawa, T.Masujima: "Real Time Analysis of Exocytosis and PKC-Translocation in RBL-2H3 Cells Visualized by Video-Enhanced Light/Fluorescence Microscopy" Neurochem.Res. in press. (1995)

K.Ozawa、T.Masujima：“通过视频增强光/荧光显微镜可视化 RBL-2H3 细胞中胞吐作用和 PKC 易位的实时分析” Neurochem.Res。

T.Masujima: "Fundamental Aspects of X-ray Photoacoustic Phenomena" J. Physique. 4-C7. 353-356 (1994)

T.Masujima：“X 射线光声现象的基本方面”J. Physique。

K.Ozawa: "Real Time Analysis of Exocytosis and PKC-translocation in RBL-2H3 Cells Visualized by Video-Enhanced Light/Fluorescence Microscopy" Neurochem. Res.20(in press). (1995)

K.Ozawa：“通过视频增强光/荧光显微镜可视化 RBL-2H3 细胞中胞吐作用和 PKC 易位的实时分析”Neurochem。

E.Kawai, E.Suzaki, K.Ozawa, T.Masujima: "The Role of Cytoskeleton in Phagocytosis-Exocytosis Sequence In a Neutrophil" Biochem.Biophys.Res.Commun. in press. (1995)

E.Kawai、E.Suzaki、K.Ozawa、T.Masujima：“细胞骨架在中性粒细胞吞噬-胞吐作用序列中的作用”Biochem.Biophys.Res.Commun。