2017Feng SprayingPlunging

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Feng, X.; Fu, Z.; Kaledhonkar, S.; Jia, Y.; Shah, B.; Jin, A.; Liu, Z.; Sun, M.; Chen, B.; Grassucci, R. A.; Ren, Y.; Jiang, H.; Frank, J. & Lin, Q. A Fast and Effective Microfluidic Spraying-Plunging Method for High-Resolution Single-Particle Cryo-EM. Structure, 2017, 25, 663-670.e3


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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