Three-dimensional reconstruction of biological macromolecular complexes from in-lens scanning electron micrographs

J Microsc. 2009 Jun;234(3):287-92. doi: 10.1111/j.1365-2818.2009.03168.x.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / chemistry
  • Actins / ultrastructure
  • Bacteriophage T4 / chemistry
  • Bacteriophage T4 / ultrastructure
  • Cryoelectron Microscopy / methods*
  • Image Processing, Computer-Assisted / methods*
  • Macromolecular Substances / chemistry*
  • Microscopy, Electron, Scanning / methods*
  • Protein Structure, Quaternary
  • Viral Tail Proteins / chemistry
  • Viral Tail Proteins / ultrastructure

Substances

  • Actins
  • Macromolecular Substances
  • Viral Tail Proteins