Three-dimensional reconstruction of biological macromolecular complexes from in-lens scanning electron micrographs.
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.
Woodward
J D
JD
Electron Microscope Unit, University of Cape Town, 7701 Cape Town, South Africa. jeremydavidwoodward@gmail.com
Wepf
R
R
Sewell
B T
BT
D000199:Actins / Q000737:chemistry / Q000648:ultrastructure; D017122:Bacteriophage T4 / Q000737:chemistry / Q000648:ultrastructure; D020285:Cryoelectron Microscopy / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D046911:Macromolecular Substances / Q000737:chemistry*; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D020836:Protein Structure, Quaternary; D017376:Viral Tail Proteins / Q000737:chemistry / Q000648:ultrastructure
D000199:Actins; D046911:Macromolecular Substances; D017376:Viral Tail Proteins