Development of X-ray-and Chemi-luminescence Video-Microscopy and its Application to the Analysis of Biodynamics in Micro Region

X射线和化学发光视频显微镜的发展及其在微区生物动力学分析中的应用

• 批准号: 04454525
• 负责人: MASUJIMA Tsutomu
• 金额: $ 3.78万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454525/
• 关键词: Chemiluminescence; X-ray luminescence; Videomicroscopy; Neutrophil; Superoxide; Cell Dynamics; シンクロトロン放射光; X線誘起発光

1)Development of Chemiluminescence Video MicroscopyBy assembly of a photon-counting video-camera and optical microscopy, photosensitive video-microscopy was developed which can be applied easily to the observation of dynamic aspects of biological systems in micro region.2)Application of this Chemiluminescence Video MicroscopyThe developed system was applied to the observation of phagocytosis of Neutrophils in which superoxide is generated to kill the foreign particles. This superoxide was detected as the chemiluminescence with reaction of luminor. The time course of this generation process and the amount of superoxide were successfully analyzed and showed future fruitful potential of this method.3)X-ray Luminescent PhenomenaIrradiating the strong X-ray from synchrotron, the intensity of luminescence was measured and the intensity depends on the hydrophobisity of solvent, and was weak in aqueous solvent. This phenomena can be applied to the monitoring of hydrophobisity of probe surrounding.4)Application of Video Analyzing System to Other Dynamic AspectsThis system was applied to the analysis of separation process of Liquid Chromatography. The band separation process in the column was directly observed by this system and the real separation process was cleared that has been only under imagination of all chromatographer.It was shown that the developed video-analyzing system has fruitful potentials for the analysis of dynamic aspects in not only micro-region but also in macro-region.

1)化学发光视频显微镜的开发通过组装光子计数摄像机和光学显微镜，开发了光敏视频显微镜，可以轻松地应用于微区域生物系统动态方面的观察。2)化学发光视频显微镜的应用开发的系统应用于中性粒细胞的吞噬作用的观察，其中产生超氧化物以杀死异物 颗粒。这种超氧化物通过与发光体反应的化学发光来检测。成功地分析了这一生成过程的时间过程和超氧化物的量，并显示了该方法未来的丰硕潜力。3）X射线发光现象在同步加速器的强X射线照射下，测量了发光强度，发光强度取决于溶剂的疏水性，在水性溶剂中发光较弱。这种现象可以应用于探针周围疏水性的监测。4)视频分析系统在其他动态方面的应用该系统应用于液相色谱分离过程的分析。该系统直接观察到色谱柱中的谱带分离过程，明确了色谱工作者只能想象的真实分离过程。结果表明，所开发的视频分析系统不仅在微观区域而且在宏观区域的动态方面具有丰富的分析潜力。

项目成果

N.Ikuta: "Searching X-ray Excited Emission Reagents in Toluene,Dioxan and Ethanol" Photon Factory Activity Report. 11(in press). (1994)

N.Ikuta：“在甲苯、二恶烷和乙醇中搜索 X 射线激发发射试剂”光子工厂活动报告。

洲崎 悦子・升島 努: "レーザー光音響法、顕微画像解析法による多項目イムノアッセイシステムの開発と微小域成分解析" 薬学雑誌. 112. 314-326 (1992)

Etsuko Suzaki 和 Tsutomu Masushima：“使用激光光声方法和显微图像分析方法开发多项目免疫测定系统和微区域成分分析”Pharmaceutical Journal 112. 314-326 (1992)。

N.Ikuta, T.Masujima, E.Kawai, H.Shiwaku, H.Ohno, M.Ando: "Searching X-ray Excited Emission Reagents in Toluene, Dioxan and Ethanol" Photon Factory Activity Report. 11(in press). (1994)

N.Ikuta、T.Masujima、E.Kawai、H.Shiwaku、H.Ohno、M.Ando：“在甲苯、二恶烷和乙醇中搜索 X 射线激发发射试剂”光子工厂活动报告。

E.Suzaki, H.Kobayashi, Y.Kodama, T.Masujima, S.Terakawa: "Video-rate Dynamics of Exocytotic Events Associated with Phagocytosis in Polymorphonuclear Leucocytes" J.Cell Biol. (in press). (1994)

E.Suzaki、H.Kobayashi、Y.Kodama、T.Masujima、S.Terakawa：“与多形核白细胞吞噬作用相关的胞吐事件的视频速率动态”J.Cell Biol。

升島 努: "好中球の異物処理機構:食胞形成、顆粒の開口放出及び活性酸素生成" ヒューマンサイエンス. 5(2)(グラビア). (1994)

Tsutomu Masushima：“中性粒细胞的异物处理机制：吞噬体形成、颗粒胞吐作用和活性氧产生”《人类科学》5(2)（凹版）。