Noble, A. J.; Wei, H.; Dandey, V. P.; Zhang, Z.; Tan, Y. Z.; Potter, C. S. and Carragher, B. Reducing effects of particle adsorption to the air-water interface in cryo-EM. Nature methods, 2018, 15, 793-795
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.