title: GraFix: sample preparation for single-particle electron cryomicroscopy.
abstract: We developed a method, named GraFix, that considerably improves sample quality for structure determination by single-particle electron cryomicroscopy (cryo-EM). GraFix uses a glycerol gradient centrifugation step in which the complexes are centrifuged into an increasing concentration of a chemical fixation reagent to prevent aggregation and to stabilize individual macromolecules. The method can be used to prepare samples for negative-stain, cryo-negative-stain and, particularly, unstained cryo-EM.
authors.affiliation: Department of Cellular Biochemsitry, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany. bkastne@gwdg.de
authors.forename: Berthold
authors.initials: B
authors.lastname: Kastner
authors.forename: Niels
authors.initials: N
authors.lastname: Fischer
authors.forename: Monika Mariola
authors.initials: MM
authors.lastname: Golas
authors.forename: Bjoern
authors.initials: B
authors.lastname: Sander
authors.forename: Prakash
authors.initials: P
authors.lastname: Dube
authors.forename: Daniel
authors.initials: D
authors.lastname: Boehringer
authors.forename: Klaus
authors.initials: K
authors.lastname: Hartmuth
authors.forename: Jochen
authors.initials: J
authors.lastname: Deckert
authors.forename: Florian
authors.initials: F
authors.lastname: Hauer
authors.forename: Elmar
authors.initials: E
authors.lastname: Wolf
authors.forename: Hannes
authors.initials: H
authors.lastname: Uchtenhagen
authors.forename: Henning
authors.initials: H
authors.lastname: Urlaub
authors.forename: Franz
authors.initials: F
authors.lastname: Herzog
authors.forename: Jan Michael
authors.initials: JM
authors.lastname: Peters
authors.forename: Dietmar
authors.initials: D
authors.lastname: Poerschke
authors.forename: Reinhard
authors.initials: R
authors.lastname: Lührmann
authors.forename: Holger
authors.initials: H
authors.lastname: Stark
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods*; D007089:Image Enhancement / Q000379:methods*; D013048:Specimen Handling / Q000379:methods*; D016707:Tissue Fixation / Q000379:methods*