title: Screening for two-dimensional crystals by transmission electron microscopy of negatively stained samples.
abstract: Structural studies of soluble and membrane proteins by electron crystallography include several critical steps. While the two-dimensional (2D) crystallization arguably may be described as the major bottleneck of electron crystallography, the screening by transmission electron microscopy (EM) to identify 2D crystals requires great care as well as practice. Both sample preparation and EM are skills that are relatively easily acquired, compared to the identification of the first ordered arrays. Added to this, membranes may have a variety of morphologies and sizes. Here we describe all steps involved in the screening for 2D crystals as well as the evaluation of samples.
authors.affiliation: School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA.
authors.forename: Tina M
authors.initials: TM
authors.lastname: Dreaden
authors.forename: Maureen
authors.initials: M
authors.lastname: Metcalfe
authors.forename: Laura Y
authors.initials: LY
authors.lastname: Kim
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Johnson
authors.forename: Bridgette A
authors.initials: BA
authors.lastname: Barry
authors.forename: Ingeborg
authors.initials: I
authors.lastname: Schmidt-Krey
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods; D003460:Crystallization / Q000379:methods*; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D016624:Negative Staining / Q000379:methods*; D011506:Proteins / Q000737:chemistry* / Q000648:ultrastructure; D011510:Proteolipids / Q000737:chemistry
chemical_list: D011506:Proteins; D011510:Proteolipids; C020485:proteoliposomes