Development of A Chromato-video Spectro-Nanoscope and Establishment of Zeptmol Separation Base

色谱视频纳米光谱仪的研制及Zeptmol分离基地的建立

• 批准号: 09554051
• 负责人: MASUJIMA Tsutomu
• 金额: $ 8.06万
• 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-09554051/
• 关键词: micro-separation technology; laser ionization; MALDI-TOF; MS; video scopes; fluorescence depolarization; gel electrophresis; チップ化分離場

1. Development of Laser Ionization & Separation Method and its EquipmentConventional matrics assisted laser desorption ionization-time of flight mass spectrometer (MALDI-TOF/MS) was first used to check the detectability of micro-amount biomolecules. The detection of histamines and insulins in a hundred cells was found to be detected and its time course and dynamics was observed. However, we found smaller variety of molecules in this detection. The reason was that minor molecules cannot be ionized due to tremendous amount of sarounding molecules which are easy to be ionized. The use of micro-bore-gel liquid chromatography priorto the mass measurement is very effective to detect various molecules altogehter.Finally, the equipment for separation by matrics assisted nitrogen laser induced ionization had been home made and the detection limit was 10 amol for bradykinine. Zept mol level detection limit can be easyly expected by using focused beam.2. Development of Molecular Movement SpectrometerThe molecular movement spectrometer was developed in colaboration with JASCO Co. Ltd., and it was applied to the antigen-antibody binding study with fluorescence depolarization detector. This equipment was very usefull to detect the molecular mobility in micro separation field of gel.3. Video Analysis of Micro-separation field with polymer matricsA nanosope for micro-separation field was also developed and it was applied for the micro-gel-separation base. The micro-sharp sample zone making was very difficult due to fast diffusion process and this point is the big obstacle for the development of chip-size integrated separation system. We discussed the future potentials of these newly developed methods in each publication.

1.激光电离分离方法及其装置的研制首次将常规基质辅助激光解吸电离飞行时间质谱仪（MALDI-TOF/MS）用于检测微量生物分子。发现在百个细胞中检测到组胺和胰岛素，并观察其时程和动力学。然而，我们在这种检测中发现了较小的分子种类。其原因是小分子不能被电离，因为大量的小分子很容易被电离。最后，自制了基质辅助氮激光诱导电离分离装置，对缓激肽的检测限为10阿莫尔mol/L。通过使用聚焦光束，可以很容易地达到Zept摩尔级检测限.分子运动光谱仪的开发分子运动光谱仪是与JASCO Co. Ltd.合作开发的，并将其应用于荧光去偏振检测器的抗原抗体结合研究。该装置对凝胶微分离领域中的分子流动性检测具有重要的意义.基于聚合物基体的微分离场视频分析还研制了一种微分离场纳米探针，并将其应用于微凝胶分离基底。由于扩散过程快，使得微小尖锐样品区的制作非常困难，这一点是芯片尺寸集成分离系统发展的一大障碍。我们讨论了这些新开发的方法在每个出版物的未来潜力。

项目成果

A.Tamura: "Direct observation of sample trapping process onto internal-surface reversed phase presample trapping process onto internal-surface reversed phase pre-column" Anal.Chim.Acta.365. 115-120 (1998)

A.Tamura：“直接观察内表面反相预柱上的样品捕获过程在内表面反相预柱上的样品捕获过程”Anal.Chim.Acta.365。

A.Tamura, M.Khademizadeh, K.Ozawa and T.Masujima: "Direct Observation of Sample Trapping Process onto Internal-Surface Reversed-phase Pre-Column" Anal.Chim.Acta.365. 115-120 (1998)

A.Tamura、M.Khademizadeh、K.Ozawa 和 T.Masujima：“内表面反相预柱上样品捕获过程的直接观察”Anal.Chim.Acta.365。

T.Masujima, K.Ozawa, A.Tamura: "Molecular Dynamic Analysis by Video Detection" Bunseki. 273(9). 720-726 (1997)

T.Masujima、K.Ozawa、A.Tamura：“通过视频检测进行分子动力学分析”Bunseki。

A.Tamura: "Direct observationof separation processes by the chromato-videoscope" J.Pharm.Biomed.Anal.15. 1477-1482 (1997)

A.Tamura：“通过色谱内窥镜直接观察分离过程”J.Pharm.Biomed.Anal.15。

A.Tamura: "Determination of Insulin Content in Pancreatic beta-Cell Line MIN6 Cells by Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry" Biol.Pharm.Bull.21・11. 1240-1241 (1998)

A.Tamura：“通过基质辅助激光解吸电离飞行时间质谱测定胰腺β细胞系MIN6细胞中的胰岛素含量”Biol.Pharm.Bull.21・11（1998）。