National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R21 GM135666
米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R01 GM126011
米国
引用
ジャーナル: Curr Opin Struct Biol / 年: 2023 タイトル: Challenges in making ideal cryo-EM samples. 著者: Bong-Gyoon Han / Agustin Avila-Sakar / Jonathan Remis / Robert M Glaeser / 要旨: Recognizing that interaction with the air-water interface (AWI) is a major challenge for cryo-EM, we first review current approaches designed to avoid it. Of these, immobilizing particles on affinity ...Recognizing that interaction with the air-water interface (AWI) is a major challenge for cryo-EM, we first review current approaches designed to avoid it. Of these, immobilizing particles on affinity grids is arguably the most promising. In addition, we review efforts to gain more reliable control of the sample thicknesses, not the least important reason being to prevent immobilized particles from coming in contact with the AWI of the remaining buffer. It is emphasized that avoiding such a contact is as important for cryo-ET as for single-particle cryo-EM. Finally, looking to the future, it is proposed that immobilized samples might be used to perform time-resolved biochemical experiments directly on EM grids rather than just in test tubes or cuvettes.