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Reduction of charging in protein electron cryomicroscopy
Author(s) -
Jacob Brink,
Heinz Gross,
Peter Tittmann,
Sherman Mb,
Wah Chiu
Publication year - 1998
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1046/j.1365-2818.1998.00342.x
Subject(s) - evaporation , materials science , cryo electron microscopy , sputtering , scanning electron microscope , electron microscope , electron diffraction , resolution (logic) , coating , cathode ray , microstructure , electrical conductor , ion , diffraction , analytical chemistry (journal) , thin film , electron , chemistry , nanotechnology , optics , composite material , chromatography , quantum mechanics , artificial intelligence , biochemistry , physics , computer science , thermodynamics , organic chemistry
Charging causes a loss of resolution in electron cryomicroscopy with biological specimens prepared without a continuous carbon support film. Thin conductive films were deposited onto catalase crystals prepared across holes using ion‐beam sputtering and thermal evaporation and evaluated for the effectiveness of charge reduction. Deposits applied by ion‐beam sputtering reduced charging but concurrently resulted in structural damage. Coatings applied by thermal evaporation also reduced charging, and preserved the specimen structure beyond 5 Å resolution as judged from electron diffraction patterns and images of glucose‐embedded catalase crystals tilted to 45° in the microscope. This study demonstrates for the first time the feasibility of obtaining high‐resolution data from unstained, unsupported protein crystals with a conductive surface coating.