title: Preservation of high resolution protein structure by cryo-electron microscopy of vitreous sections.
abstract: We have quantitated the degree of structural preservation in cryo-sections of a vitrified biological specimen. Previous studies have used sections of periodic specimens to assess the resolution present, but preservation before sectioning was not assessed and so the damage due particularly to cutting was not clear. In this study large single crystals of lysozyme were vitrified and from these X-ray diffraction patterns extending to better than 2.1A were obtained. The crystals were high pressure frozen in 30% dextran, and cryo-sectioned using a diamond knife. In the best case, preservation to a resolution of 7.9A was shown by electron diffraction, the first observation of sub-nanometre structural preservation in a vitreous section.
authors.affiliation: Institute of Molecular and Cellular Biology, University of Leeds, LS2 9JT, UK. k.sader@leeds.ac.uk
authors.forename: Kasim
authors.initials: K
authors.lastname: Sader
authors.forename: Daniel
authors.initials: D
authors.lastname: Studer
authors.forename: Benoît
authors.initials: B
authors.lastname: Zuber
authors.forename: Helmut
authors.initials: H
authors.lastname: Gnaegi
authors.forename: John
authors.initials: J
authors.lastname: Trinick
mesh_terms: D000818:Animals; D002645:Chickens; D020285:Cryoelectron Microscopy / Q000379:methods*; D015925:Cryopreservation / Q000379:methods*; D016616:Cryoultramicrotomy / Q000379:methods*; D003460:Crystallization; D004529:Egg White; D005260:Female; D009113:Muramidase / Q000737:chemistry / Q000648:ultrastructure; D014961:X-Ray Diffraction
chemical_list: D009113:Muramidase