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

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

Feng, Xiangsong;Fu, Ziao;Kaledhonkar, Sandip;Jia, Yuan;Shah, Binita;Jin, Amy;Liu, Zheng;Sun, Ming;Chen, Bo;Grassucci, Robert A;Ren, Yukun;Jiang, Hongyuan;Frank, Joachim;Lin, Qiao;
structure (london, england : 1993) 2017 Vol. 25 pp. 663-670.e3
213
feng2017astructure

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.

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