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

作者：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^*

来源：Structure, 2017, 25(4): 663-+.

DOI：10.1016/j.str.2017.02.005

• ELECTRON CRYOMICROSCOPY
• VITRIFIED ICE
• LIQUID JET
• MICROSCOPY
• ACQUISITION
• MECHANISM
• RIBOSOME
• RECONSTRUCTION
• ATOMIZATION
• MICROGRAPHS

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.

• 出版日期2017-4-4
• 单位哈尔滨工业大学