title: MicroED data collection and processing.
abstract: MicroED, a method at the intersection of X-ray crystallography and electron cryo-microscopy, has rapidly progressed by exploiting advances in both fields and has already been successfully employed to determine the atomic structures of several proteins from sub-micron-sized, three-dimensional crystals. A major limiting factor in X-ray crystallography is the requirement for large and well ordered crystals. By permitting electron diffraction patterns to be collected from much smaller crystals, or even single well ordered domains of large crystals composed of several small mosaic blocks, MicroED has the potential to overcome the limiting size requirement and enable structural studies on difficult-to-crystallize samples. This communication details the steps for sample preparation, data collection and reduction necessary to obtain refined, high-resolution, three-dimensional models by MicroED, and presents some of its unique challenges.
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.
authors.forename: Johan
authors.initials: J
authors.lastname: Hattne
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.
authors.forename: Francis E
authors.initials: FE
authors.lastname: Reyes
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.
authors.forename: Brent L
authors.initials: BL
authors.lastname: Nannenga
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.
authors.forename: Dan
authors.initials: D
authors.lastname: Shi
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.
authors.forename: M Jason
authors.initials: MJ
authors.lastname: de la Cruz
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge, England.
authors.forename: Andrew G W
authors.initials: AG
authors.lastname: Leslie
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.
authors.forename: Tamir
authors.initials: T
authors.lastname: Gonen
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods*; D018360:Crystallography, X-Ray / Q000379:methods*; D003625:Data Collection / Q000379:methods*; D011506:Proteins / Q000737:chemistry*
chemical_list: D011506:Proteins
keywords: MicroED; cryo-EM; crystallography; electron diffraction; nanocrystals