Observation and analysis of muscle tissue by high-speed AFM

2018 Fiscal Year Final Research Report

• Project/Area Number: 17K15149
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Morphology/Structure
• Research Institution: Nagoya University
• Principal Investigator: Usukura Eiji 名古屋大学, 未来材料システム研究所, 研究員 (00643727)
• Research Collaborator: Usukura Jiro ; Narita Akihiro
• Project Period (FY): 2017-04-01 – 2019-03-31
• Keywords: 高速AFM / 徳安法 / 組織 / 細胞 / 蛍光顕微鏡

Outline of Final Research Achievements

We succeeded the observation of the sarcomere and the photoreceptor cell by the combination high speed AFM and Tokuyasu method improved for AFM. Moreover, the correlation observation between high speed AFM and fluorescence microscope was established by using immunocytochemistry.
In case of observation of sarcomere, M line and Z disk were confirmed clearly. Additionally, we thought that there were myosin head in the myosin filament and myomesin in the M line. On the other hand, specific structures of outer and inner segment were confirmed in case of photoreceptor cell. Furthermore, we tried to observe the rhodopsin with antibody staining for correlation between high speed AFM and fluorescence microscope. Interestingly, we detected many complexes by coupling a primary antibody and a secondary antibody on the outer segment.

Free Research Field

生物物理

Academic Significance and Societal Importance of the Research Achievements

本研究で行った徳安法と高速AFMを用いたサルコメアと視細胞の観察では、これまでに電子顕顕微鏡でしか踏み込めなかった組織切片の高分解能観察を実現できた。しかも、金属蒸着や高真空環境は必要とせず、生体分子本来の水環境条件下で観察を行えたことは重要である。そして、この観察手法は他の組織にも応用することができる。さらに、免疫細胞化学と組み合わせることで、高速AFMと蛍光顕微鏡の相関観察の可能性もしめすことができた。