Tan, Y. Z.; Baldwin, P. R.; Davis, J. H.; Williamson, J. R.; Potter, C. S.; Carragher, B. & Lyumkis, D. Addressing preferred specimen orientation in single-particle cryo-EM through tilting. Nature methods, 2017, 14, 793-796
We present a strategy for tackling preferred specimen orientation in single-particle cryogenic electron microscopy by employing tilts during data collection. We also describe a tool to quantify the resulting directional resolution using 3D Fourier shell correlation volumes. We applied these methods to determine the structures at near-atomic resolution of the influenza hemagglutinin trimer, which adopts a highly preferred specimen orientation, and of ribosomal biogenesis intermediates, which adopt moderately preferred orientations.