Establishment of Post-Proteomic Analysis by the Development of Video-Mass-Scope Method

通过视频质谱法的发展建立后蛋白质组分析

• 批准号: 13470479
• 负责人: MASUJIMA Tsutomu
• 金额: $ 9.54万
• 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: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470479/
• 关键词: Video-microscope; Mass Spectrometer; Video-Mass-Scope; Single Cell Mass Spectrometry; Single Cell Analysis; Cell Molecular Dynamism; Image Analysis; ビデオマイクロスコープ; ポストプロテーオーム解析; ビデオ-マススコープ; 可視化解析; プロテオーム; ポストゲノム

1) Development of Sensitive Mass Spectrometer : In order to analyze the secreted molecules from a cell, an ion-trap-TOF mass spectrometer and a TOP mass spectrometer with Nano-spray ionization source were developed.2) Development of Molecular Trapping Interface : Nano-spray type ion trapping tips were investigated and the tip in which pre-concentrating resins were embedded inside, was developed and applied to pre-concentration of secreted molecules from Mast cells.3) Development of the Video-Mass-Scope : By combination of the equipments 1. and method 2., the video-mass-scope was developed and applied to the detection of secreted molecules from the cells. On the stimulation of Mast Cells, Histamine and Serotonin were successfully detected by this method. This research was presented in the 123-th Annual Meetings of Japan Pharmaceutical Society.4) Single Cell MALDI-TOF Mass Spectrometric Analysis : This method was developed and successfully applied to analyze the increase of Histamine contents in a single Mast Cell according to the cell maturation process and the report was published.5) Development of Pin Fiber Video-Micro-Scope : This scope was also developed in order to investigate a single cell for long time with little irradiation damage to the cell. This method is also found to be useful to apply to the single molecular detection.The fundamentals and related methodologies were successfully developed by this study and the potentials of these methods were high enough to the direct and simultaneous observation of cell behavior and its dynamic changes in molecular contents.

1)灵敏质谱仪的研制：为了分析细胞分泌的分子，研制了离子阱-TOF质谱仪和带有纳米喷雾电离源的TOP质谱仪。2）分子捕获接口的研制：研究了纳米喷雾型离子捕集针尖，3）视频质谱仪的研制：通过设备的组合1.以及方法2，开发了视频质谱仪并将其应用于检测细胞分泌的分子。在肥大细胞刺激下，用该方法成功地检测了组胺和5-羟色胺。该研究在日本药学会第123届年会上发表。4）单细胞MALDI-TOF质谱分析：该方法已开发并成功应用于根据细胞成熟过程分析单个肥大细胞中组胺含量的增加并发表报告。5）Pin Fiber Video-Micro-Scope的开发：该范围也是为了长时间研究单个细胞而对细胞的辐射损伤很小而开发的。本研究为细胞行为及其分子含量的动态变化的直接、同步观测奠定了基础，并为细胞行为的实时监测奠定了基础。

项目成果

M.Suzuzto, Y.Hirakawa, T.Shingaki, T.Masujima 他: "Nano-Kinetics of Probe-Particles in Solution Visualized by a Pin-Fiber Video Scope"Analytical Sciences. 19. 43-47 (2003)

M.Suzuzto、Y.Hirakawa、T.Shingaki、T.Masujima 等人：“通过针光纤视频示波器可视化溶液中探针粒子的纳米动力学”分析科学 19. 43-47 (2003)。

M.Shimizu, T.Shingaki, Y.Hirakawa, T.Masujima 他: "Development of the Single-Cell MALDI-TOF(Matrix-Assisted Laser Desorption/lonization Time-of-Flight) Mass-Spectroscopic Assay"Analytical Sciences. 19. 49-53 (2003)

M.Shimizu、T.Shingaki、Y.Hirakawa、T.Masujima 等人：“单细胞 MALDI-TOF（基质辅助激光解吸/电离飞行时间）质谱分析的开发”分析科学. 19 .49-53 (2003)

N. Ojima, T. Shingaki, T. Yamamoto, T. Masujima: "Droplet Electro-coupling between capillary electrophoresis and matrix assisted laser desorption ionization time of flight mass spectroscopy and its application"Electrophorelsis. 22. 3478-3482 (2001)

N. Ojima、T. Shiningaki、T. Yamamoto、T. Masujima：“毛细管电泳与基质辅助激光解吸电离飞行时间质谱之间的液滴电耦合及其应用”电泳。

T. Masujima, N. Ojima, T. Tuda Ed.: "Scientific Experiments II -- Guarding life from invader --"Maruzen. 160 (2001)

T. Masujima、N. Ojima、T. Tuda Ed.：《科学实验 II——保护生命免受入侵者——》丸善。

升島努, 尾島典行: "楽しい科学実験 II 試みから想像へ ----侵入者から命を守る----"丸善株式会社. 160 (2001)

增岛勉、小岛典之：《有趣的科学实验 II：从尝试到想象----保护生命免受入侵者侵害----》丸善株式会社 160 (2001)