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. Epub 2018 Sep 24.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Air*
  • Apoferritins / ultrastructure*
  • Cryoelectron Microscopy / methods*
  • Hemagglutinins / ultrastructure*
  • Humans
  • Receptor, Insulin / ultrastructure*
  • Water / chemistry*

Substances

  • Hemagglutinins
  • Water
  • Apoferritins
  • Receptor, Insulin