title: Cryomicroscopy of radiation sensitive specimens on unmodified graphene sheets: reduction of electron-optical effects of charging.
abstract: Images of radiation-sensitive specimens obtained by electron microscopy suffer a reduction in quality beyond that expected from radiation damage alone due to electron beam-induced charging or movement of the specimen. For biological specimens, charging and movement are most severe when they are suspended in an insulating layer of vitreous ice, which is otherwise optimal for preserving hydrated specimens in a near native state. We image biological specimens, including a single particle protein complex and a lipid-enveloped virus in thin, vitreous ice films over suspended sheets of unmodified graphene. We show that in such preparations, the charging of ice, as assessed by electron-optical perturbation of the imaging beam, is eliminated. We also use the same specimen supports to record high resolution images at liquid nitrogen temperature of monolayer paraffin crystals grown over graphene.
authors.affiliation: Division of Physical Biochemistry, MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, United Kingdom. Electronic address: kasim.sader@gmail.com.
authors.forename: Kasim
authors.initials: K
authors.lastname: Sader
authors.affiliation: Division of Physical Biochemistry, MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, United Kingdom.
authors.forename: Martyn
authors.initials: M
authors.lastname: Stopps
authors.affiliation: Division of Physical Biochemistry, MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, United Kingdom.
authors.forename: Lesley J
authors.initials: LJ
authors.lastname: Calder
authors.affiliation: Division of Physical Biochemistry, MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, United Kingdom. Electronic address: prosent@nimr.mrc.ac.uk.
authors.forename: Peter B
authors.initials: PB
authors.lastname: Rosenthal
mesh_terms: D001052:Apoferritins / Q000648:ultrastructure; D020285:Cryoelectron Microscopy / Q000379:methods*; D015925:Cryopreservation / Q000379:methods*; D003461:Crystallography; D006108:Graphite / Q000737:chemistry*; D046529:Microscopy, Electron, Transmission / Q000379:methods; D009975:Orthomyxoviridae / Q000648:ultrastructure; D010232:Paraffin / Q000737:chemistry; D014771:Virion / Q000648:ultrastructure
chemical_list: D006108:Graphite; D010232:Paraffin; D001052:Apoferritins
keywords: Beam induced specimen movement; Charging; Cryomicroscopy; Graphene; Radiation damage