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Time-resolved cryo-EM using Spotiton.
Nature Methods ( IF 36.1 ) Pub Date : 2020-08-10 , DOI: 10.1038/s41592-020-0925-6
Venkata P Dandey 1 , William C Budell 1 , Hui Wei 1 , Daija Bobe 1 , Kashyap Maruthi 1 , Mykhailo Kopylov 1 , Edward T Eng 1 , Peter A Kahn 2 , Jenny E Hinshaw 3 , Nidhi Kundu 3 , Crina M Nimigean 4 , Chen Fan 4 , Nattakan Sukomon 4 , Seth A Darst 5 , Ruth M Saecker 5 , James Chen 5 , Brandon Malone 5 , Clinton S Potter 1, 6 , Bridget Carragher 1, 6
Affiliation  

We present an approach for preparing cryo-electron microscopy (cryo-EM) grids to study short-lived molecular states. Using piezoelectric dispensing, two independent streams of ~50-pl droplets of sample are deposited within 10 ms of each other onto the surface of a nanowire EM grid, and the mixing reaction stops when the grid is vitrified in liquid ethane ~100 ms later. We demonstrate this approach for four biological systems where short-lived states are of high interest.



中文翻译:

使用 Spotiton 的时间分辨冷冻电镜。

我们提出了一种制备低温电子显微镜 (cryo-EM) 网格以研究短寿命分子状态的方法。使用压电分配,两个独立的 ~50 pl 样品液滴流在 10 毫秒内沉积到纳米线 EM 网格的表面上,当网格在液体乙烷中玻璃化约 100 毫秒后,混合反应停止。我们为四种生物系统展示了这种方法,其中短命状态非常受关注。

更新日期:2020-08-10
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