Nanoanalysis of the molecular mechanism of chromatin functions using single molecule imaging in vio and super-resolution microscopy

2017 Fiscal Year Final Research Report

• Project Area: Dynamic chromatin structure and function
• Project/Area Number: 25116007
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: Tokunaga Makio 東京工業大学, 生命理工学院, 教授 (00192659)
• Co-Investigator(Renkei-kenkyūsha): SOGAWA KUMIKO (SAKATA KUMIKO) 東北大学, 大学院・農学研究科・学術研究院 (20291073) ; ITO YUMA 東京工業大学, 生命理工学院, 助教 (70803245)
• Project Period (FY): 2013-06-28 – 2018-03-31
• Keywords: 生物物理 / 超解像イメージング / １分子イメージング / 細胞情報・動態 / クロマチン動構造 / ナノ定量解析 / 生体分子計測 / 生細胞分子動態

Outline of Final Research Achievements

For the purpose of understanding the molecular mechanism of chromatin functions, we have developed integrated microscopy with improved resolution in multi-color single-molecule super-resolution microscopy by optimizing illumination and stabilization of optics. We have also developed a new method, moving subtrajectory analysis using single-molecule tracking, which provides us spatiotemporal to quantification of not only dynamics but also the kinetics of interactions using single-color images. Combined use of this with multi-color simultaneous imaging and super-resolution microscopy, enables integrated analysis of different kinds of molecules with a spatiotemporal resolution and in relation to cellular activity. Using these techniques, we carried out imaging analysis of molecular dynamics and interactions of nucleosomes, RNA polymerase II, histone variants, transcription regulatory proteins, the INO80 chromatin remodeling complex, and the multilayer structure of the nucleosome.

Free Research Field

生物物理学