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

1. 激光电离分离方法及其设备的发展传统材料辅助激光解吸电离飞行时间质谱仪（MALDI-TOF/MS）首次用于检测微量生物分子的可检出性。在100个细胞中检测到组胺和胰岛素，并观察其时间过程和动态。然而，我们在这次检测中发现的分子种类较少。原因是小分子不能被电离，因为周围有大量容易被电离的分子。在质量测量之前使用微孔-凝胶液相色谱法可以有效地同时检测各种分子。自制了基质辅助氮激光诱导电离分离装置，对缓激碱的检出限为10amol。使用聚焦光束可以很容易地预测Zept的mol级检测限。分子运动光谱仪的发展分子运动光谱仪是与日本JASCO有限公司合作开发的，并应用于荧光去极化检测器的抗原抗体结合研究。该仪器可用于凝胶微分离领域中分子迁移率的检测。开发了用于微分离场的纳米显微镜，并将其应用于微凝胶分离基质。由于扩散过程快，微尖样品区制造非常困难，这是芯片级集成分离系统发展的一大障碍。我们在每篇文章中讨论了这些新开发方法的未来潜力。

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.