Establishment of observation technique for living cells with both photo excitation and chemiexcitation

2015 Fiscal Year Final Research Report

• Project/Area Number: 25440162
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Morphology/Structure
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: Hideto Hoshino 国立研究開発法人産業技術総合研究所, バイオメディカル研究部門, 主任研究員 (20371073)
• Co-Investigator(Renkei-kenkyūsha): UEGAKI Koichi 国立研究開発法人産業技術総合研究所, バイオメディカル研究部門, 研究グループ長 (00356544) ; NIWA Kazuki 国立研究開発法人産業技術総合研究所, 計量標準総合センター物理計測標準部門, 主任研究員 (30443211)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: 閉鎖型灌流培養観察系 / 化学発光 / 生細胞観察

Outline of Final Research Achievements

Making full use of a conventional fluorescent microscope system, we tried chemiluminescent live-cell imaging for the stably expressing cell lines which we introduced the BAF (BRET-based Auto-illuminated Fluorescent-protein)-fusion proteins that are our own technique into, and showed the possibility of the chemiluminescent live-cell imaging based on luciferin - luciferase reaction. In the experiments, we found some problems newly. A major problem was the air bubbles dependent on the required culture medium ingredient and caused by a perfusion process in the closed live-cell micro-observation chamber system. We tried the development of the device to remove the air bubbles based on the simple principle concerned, but did not lead to completion during the research period. Meanwhile, for the purpose of a higher intensity of eBAF-Y, we found a new approach to achieve a higher efficiency of the resonance energy transfer which happens in the BAF molecules, and applied for a Japanese patent.

Free Research Field

細胞生物学