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

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.

SUBMITTER: Feng X 

PROVIDER: S-EPMC5382802 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature

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

Feng Xiangsong X   Fu Ziao Z   Kaledhonkar Sandip S   Jia Yuan Y   Shah Binita B   Jin Amy A   Liu Zheng Z   Sun Ming M   Chen Bo B   Grassucci Robert A RA   Ren Yukun Y   Jiang Hongyuan H   Frank Joachim J   Lin Qiao Q  

Structure (London, England : 1993) 20170309 4

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 i  ...[more]

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