Development of fluorescence labeling methods of proteins in cells and tissues for high-resolution correlated light-electron microscopy (CLEM) using Atmospheric scanning electron microscopy

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K14499
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biophysics
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: SATO Chikara 国立研究開発法人産業技術総合研究所, 生命工学領域, 研究グループ長 (00357146)
• Co-Investigator(Kenkyū-buntansha): 海老原 達彦 国立研究開発法人産業技術総合研究所, 生命工学領域, 研究グループ付 (00344119) ; 岡田 知子 国立研究開発法人産業技術総合研究所, その他部局等, 研究員 (30344146)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 電子顕微鏡 / バイオイメージング / 電子線励起 / 光電子相関顕微鏡 / 転移性がん細胞

Outline of Final Research Achievements

Proteins associate with other proteins and form protein complexes, localizing in a specific positions in cells and tissues for their functions. The correlative light-electron microscopy (CLEM) of a sample in liquid is highly required to determine their localization at high resolution. To realize electron microscopy in solution, environmental-capsule electron microscopy (EC-EM) has been developed. However, the limited space around the sample holder of standard TEM, preclude the simultaneous observation using OM with high resolution objective lens of large NA. We have developed atmospheric scanning electron microscopy (ASEM) with inverted SEM to allow the use of an open sample container sandwiched by OM and SEM. In this research, we have tested many fluorescent substances, including GFP, qDot and ZnO. Further the markers tested here should be exploited for the application to various biological fields including basic biology, medical research, agriculture, drug discovery and diagnosis.

Free Research Field

タンパク質のクライオ電子顕微鏡法開発と電顕・光顕相関同時観察法開発による細胞・組織のタンパク局在研究