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Three‐dimensional reconstruction of biological macromolecular complexes from in‐lens scanning electron micrographs
Author(s) -
WOODWARD J.D.,
WEPF R.,
SEWELL B.T.
Publication year - 2009
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 - 10.1111/j.1365-2818.2009.03168.x
Subject(s) - micrograph , lens (geology) , optics , single particle analysis , electron microscope , scanning electron microscope , microscopy , electron micrographs , scanning transmission electron microscopy , materials science , cryo electron microscopy , electron , macromolecule , electron tomography , scanning confocal electron microscopy , particle (ecology) , field emission gun , chemistry , physics , nuclear magnetic resonance , biology , biochemistry , aerosol , organic chemistry , quantum mechanics , ecology
Summary Two helical samples: F‐actin and the bacteriophage T4 tail sheath were reconstructed in three dimensions from contrast enhanced (rotational shadowing and negatively stained) in‐lens cryo‐field emission scanning electron micrographs, using the iterative real‐space helical reconstruction method. The F‐actin – and bacteriophage T4 reconstructions compare favourably to an atomic model refined against fibre diffraction data and a cryo‐electron microscopy reconstruction, respectively. These results show that single‐particle methods, developed for macromolecules imaged in the transmission electron microscope can be applied to cryo‐field emission scanning electron micrographs data with appropriate symmetry.