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[Structure] A Fast and Effective Microfluidic Spraying-Plunging Method for High-Resolution Single-Pa


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.

Xiangsong Feng1, 2, 8, Ziao Fu3, 8, Sandip Kaledhonkar4, Yuan Jia1, Binita Shah5, Amy Jin4, 10, Zheng Liu4, Ming Sun6, Bo Chen6, 9, Robert A. Grassucci4, 7, Yukun Ren2, Hongyuan Jiang2, Joachim Frank4, 6, 7, 11, Qiao Lin1, , 1 Department of Mechanical Engineering, Columbia University, New York, NY 10027, USA 2 School of Mechatronics Engineering, Harbin Institute of Technology, Harbin 150001, China 3 Integrated Program in Cellular, Molecular, and Biophysical Studies, Columbia University College of Physicians and Surgeons, New York, NY 10032, USA 4 Department of Biochemistry and Molecular Biophysics, Columbia University New York, NY 10027, USA 5 Department of Biological Sciences, Barnard College, New York, NY 10027, USA 6 Department of Biological Sciences, Columbia University, New York, NY 10027, USA 7 Howard Hughes Medical Institute, Columbia University, New York, NY 10032, USA

http://dx.doi.org/10.1016/j.str.2017.02.005

Link:

http://www.sciencedirect.com/science/article/pii/S0969212617300369

#031317 #cryoem #spraying #plunging #highresolution

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