Reducing effects of particle adsorption to the air-water interface in cryo-EM

Nat Methods. 2018 Oct;15(10):793-795. doi: 10.1038/s41592-018-0139-3. | PubMed

Alex J Noble¹, Hui Wei¹, Venkata P Dandey¹, Zhening Zhang¹, Yong Zi Tan^1,2, Clinton S Potter^1,2, Bridget Carragher^3,4

1. National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY, USA.
2. Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, USA.
3. National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY, USA. bcarr@nysbc.org.
4. Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, USA. bcarr@nysbc.org.

Abstract

Most protein particles prepared in vitreous ice for single-particle cryo-electron microscopy (cryo-EM) are adsorbed to air-water or substrate-water interfaces, which can cause the particles to adopt preferred orientations. By using a rapid plunge-freezing robot and nanowire grids, we were able to reduce some of the deleterious effects of the air-water interface by decreasing the dwell time of particles in thin liquid films. We demonstrated this by using single-particle cryo-EM and cryo-electron tomography (cryo-ET) to examine hemagglutinin, insulin receptor complex, and apoferritin.

Presented By Alex Noble