Challenges in making ideal cryo-EM samples.
Challenges in making ideal cryo-EM samples.
复制标题
制作理想冷冻电镜样品的挑战。
DOI：
10.1016/j.sbi.2023.102646
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发表时间：
2023
影响因子：
6.8
通讯作者：
Glaeser,RobertM
中科院分区：
文献类型：
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作者：
Han,Bong-Gyoon;Avila-Sakar,Agustin;Remis,Jonathan;Glaeser,RobertM
认识到与空气-水界面（AWI）的相互作用是cryo-EM的一个主要挑战，我们首先回顾了目前设计的方法，以避免它。其中，亲和网格上的固定粒子可以说是最有前途的。此外，我们审查的努力，以获得更可靠的控制样品的厚度，而不是最不重要的原因是，以防止固定化的颗粒与AWI的剩余缓冲液接触。它强调，避免这种接触是重要的冷冻ET为单粒子冷冻EM。最后，展望未来，有人提出，固定化的样品可能会被用来执行时间分辨的生化实验直接在EM网格，而不仅仅是在试管或比色皿。
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.