Establishment of technique for supersaturation control in liquid cell transmission electron microscopy and its application to protein crystallization

2022 Fiscal Year Final Research Report

• Project/Area Number: 20H02580
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 28040:Nanobioscience-related
• Research Institution: Hokkaido University
• Principal Investigator: Yamazaki Tomoya 北海道大学, 低温科学研究所, 特任助教 (50735032)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: 透過型電子顕微鏡 / 溶液セル / タンパク質 / 過飽和度 / 誘電泳動 / 試料冷却TEMホルダー / 放射線分解 / 結晶化

Outline of Final Research Achievements

To elucidate the crystallization process of protein crystals, it is necessary to observe the crystallization process with high temporal and spatial resolution. Such observation can be performed with a liquid-cell transmission electron microscopy. In order to perform efficient crystallization in the liquid cell, techniques to control supersaturation in the liquid cell were investigated. Three techniques were developed: radiolysis combined with anti-solvent, dielectrophoresis; specimen cooling. The application of these techniques to protein crystallization was investigated.

Free Research Field

結晶成長

Academic Significance and Societal Importance of the Research Achievements

結晶化の物理・化学的メカニズムの解明は長年に渡り取り組まれている学術的に重要な課題である。また結晶化は、創薬に関わるタンパク質分子の構造解析、産業に関わる半導体などの材料開発、地球の熱収支に関わる雲の生成などの基礎であり、社会的に重要な現象である。これを調べるのに有効な溶液セル透過型電子顕微鏡法において、結晶化を能動的に生じさせる手法を開発した。これにより結晶化の瞬間を捉えることが容易になり、結晶化の理解が飛躍的に進展することが期待できる。