Development of A Fiber Nano-scope with Molecular Trapping and its Application to the Analysis of Molecular Mechanism of Allergy

分子捕获纤维纳米镜的研制及其在过敏分子机制分析中的应用

• 批准号: 09470495
• 负责人: MASUJIMA Tsutomu
• 金额: $ 8.64万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470495/
• 关键词: Molecular Trapping; HPLC resin; MALDI-TOF Mass Spectrometry; Video Nanoscope; Optical Fiber; Cell Dynamism; Brownian Motion

1. Development of Molecular Trapping Method and its Application to the Analysis of AllergySecreted molecules out of a mast cell model, RBL-2H3, were trapped by ion-exchange resins for HPLC in a single cell revel and its time course of secretion was monitored and analyzed by a fiber nano-scope. One species of the trapped molecules are identified to be histamine by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS). The molecular trapping probe of fiber type with ODS residues was also developed.2. Development of A Fiber Nano-scopeA video-nanoscope with micro spot lighting with optical fiber light-guide was also developed. The Brownian movement of colloidal gold nano-particles was observed by this video scope and the movement of particleswas analyzed to evaluate the diameters of the particles. The dynamic movement of bound gold-labeled molecules was also observed by this new nano-scope.3. Molecular Trapping and MALDI-TOF/MS AnalysisNot only the histamine but also the insulin which was secreted out of the beta pancreatic model cells was found to be detected and applied to the time course analysis of the beta pancreatic cell response.4. Application to Another SystemThis method was also applied to the analysis of phagocytosis of neutrophils to clarify the sequential dynamism of the cells and clarified many new aspects of its behavior.

1.分子捕获方法的发展及其在过敏分析中的应用使用离子交换树脂捕获肥大细胞模型RBL-2H3中的分泌分子，用于单细胞水平的HPLC，并通过纤维纳米镜监测和分析其分泌的时间过程。通过基质辅助激光解吸电离飞行时间质谱 (MALDI-TOF/MS)，被捕获的分子中的一种被鉴定为组胺。研制了ODS残留纤维型分子捕获探针。 2．光纤纳米镜的开发还开发了带有光纤光导的微点照明的视频纳米镜。通过该视频示波器观察胶体金纳米颗粒的布朗运动，并分析颗粒的运动以评估颗粒的直径。这种新型纳米显微镜还可以观察到结合的金标记分子的动态运动。3．分子捕获和MALDI-TOF/MS分析不仅检测到了β胰腺模型细胞分泌的组胺，还检测到了胰岛素，并将其应用于β胰腺细胞反应的时程分析。4．应用于另一个系统该方法还应用于分析中性粒细胞的吞噬作用，以阐明细胞的连续动态并阐明其行为的许多新方面。

项目成果

K.Ozawa: "Simultaneous analysis of membrane potential and calcium mobilization in a pancreatic β-cell line MIN6 by use of a double-probe imaging microscope system" Anal.Chim.Acta.365. 3-7 (1998)

K.Ozawa：“使用双探针成像显微镜系统同时分析胰腺 β 细胞系 MIN6 中的膜电位和钙动员”Anal.Chim.Acta.365 (1998)。

A.Tamura, T.Azuma, K.Ikeda, K.Ozawa, T.Masujima: "Determination of Insulin Content in Pancreatic beta-Cell Line MIN6 Cells aby Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry" Biol.Pharm.Bull.21(11). 1240-1241 (1998)

A.Tamura、T.Azuma、K.Ikeda、K.Ozawa、T.Masujima：“通过基质辅助激光解吸电离飞行时间质谱法测定胰腺 β 细胞系 MIN6 细胞中的胰岛素含量”Biol。

E.Suzaki: "Video-rate Dynamics of Exocytotic Events Associated With Phagocytosis in Neutrophils" Cell Motility and the Cytoskeleton. 38. 215-228 (1997)

E.Suzaki：“与中性粒细胞吞噬作用相关的胞吐事件的视频速率动力学”细胞运动和细胞骨架。

M.Suzuto, A.Nakamura, Y.Ishihara, K.Ikeda, A.Tamura, T.Masujima: "Evaluation of Diameter of Colloidal Golds by Video-Analysis of its Brownian Motions" Anal.Chim.Acta. (in press). (1999)

M.Suzuto、A.Nakamura、Y.Ishihara、K.Ikeda、A.Tamura、T.Masujima：“通过视频分析布朗运动来评估胶体金的直径”Anal.Chim.Acta。

T.Masujima: "Trapping of the secreted molecules from a single cell using HPLC resin particles under video-microscope" Biol.Pharm.Bull.21・8. 886-888 (1998)

T.Masujima：“在视频显微镜下使用 HPLC 树脂颗粒捕获单个细胞的分泌分子”Biol.Pharm.Bull.21・8（1998）。