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

1.分子捕获法的建立及其在变态反应分析中的应用用离子交换树脂对肥大细胞模型RBL-2 H3的分泌分子进行单细胞捕获，并用纤维纳米显微镜监测和分析其分泌的时间过程。基质辅助激光解吸电离-飞行时间质谱（MALDI-TOF/MS）鉴定了其中一种被捕获的分子是组胺。并研制了ODS残留纤维型分子捕获探针.光纤纳米显微镜的研制还研制了一种光纤导光微光斑照明的视频纳米显微镜。利用该视频显微镜观察了胶体金纳米颗粒的布朗运动，并对颗粒的运动进行了分析，以评价颗粒的粒径。利用这种新型的纳米显微镜还观察到了结合金标记分子的动态运动。分子捕获和MALDI-TOF/MS分析不仅检测到β胰腺模型细胞分泌的组胺，还检测到胰岛素，并应用于β胰腺细胞反应的时程分析.应用于另一个系统这种方法也被应用于中性粒细胞的吞噬作用的分析，以澄清细胞的顺序动力学，并澄清其行为的许多新方面。

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.