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Literature DB >> 28286002

A Fast and Effective Microfluidic Spraying-Plunging Method for High-Resolution Single-Particle Cryo-EM.

Xiangsong Feng¹, Ziao Fu², Sandip Kaledhonkar³, Yuan Jia⁴, Binita Shah⁵, Amy Jin³, Zheng Liu³, Ming Sun⁶, Bo Chen⁶, Robert A Grassucci⁷, Yukun Ren⁸, Hongyuan Jiang⁸, Joachim Frank⁹, Qiao Lin¹⁰.

Abstract

We describe a spraying-plunging method for preparing cryoelectron microscopy (cryo-EM) grids with vitreous ice of controllable, highly consistent thickness using a microfluidic device. The new polydimethylsiloxane (PDMS)-based sprayer was tested with apoferritin. We demonstrate that the structure can be solved to high resolution with this method of sample preparation. Besides replacing the conventional pipetting-blotting-plunging method, one of many potential applications of the new sprayer is in time-resolved cryo-EM, as part of a PDMS-based microfluidic reaction channel to study short-lived intermediates on the timescale of 10-1,000 ms. Published by Elsevier Ltd.

Entities: Chemical

Keywords: EM grid preparation; PDMS-based microsprayer; apoferritin; ice thickness; time-resolved cryo-EM

Mesh：

Substances：

Year: 2017 PMID： 28286002 PMCID： PMC5382802 DOI： 10.1016/j.str.2017.02.005

Source DB: PubMed Journal: Structure ISSN： 0969-2126 Impact factor: 5.006

44 in total

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41 in total

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Authors: Venkata P Dandey; Hui Wei; Alex J Noble; Julia Brasch; Jillian Chase; Priyamvada Acharya; Yong Zi Tan; Zhening Zhang; Laura Y Kim; Giovanna Scapin; Micah Rapp; Edward T Eng; William J Rice; Anchi Cheng; Carl J Negro; Lawrence Shapiro; Peter D Kwong; David Jeruzalmi; Amedee des Georges; Clinton S Potter; Bridget Carragher
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Authors: Tina Uroda; Isabel Chillón; Paolo Annibale; Jean-Marie Teulon; Ombeline Pessey; Manikandan Karuppasamy; Jean-Luc Pellequer; Marco Marcia
Journal: Nat Protoc Date: 2020-05-25 Impact factor: 13.491