Achievable resolution from images of biological specimens acquired from a 4k x 4k CCD camera in a 300-kV electron cryomicroscope

J Struct Biol. 2008 Jul;163(1):45-52. doi: 10.1016/j.jsb.2008.04.001. Epub 2008 Apr 14.

Abstract

Bacteriorhodopsin and epsilon 15 bacteriophage were used as biological test specimens to evaluate the potential structural resolution with images captured from a 4k x 4k charge-coupled device (CCD) camera in a 300-kV electron cryomicroscope. The phase residuals computed from the bacteriorhodopsin CCD images taken at 84,000x effective magnification averaged 15.7 degrees out to 5.8-A resolution relative to Henderson's published values. Using a single-particle reconstruction technique, we obtained an 8.2-A icosahedral structure of epsilon 15 bacteriophage with the CCD images collected at an effective magnification of 56,000x. These results demonstrate that it is feasible to retrieve biological structures to a resolution close to 2/3 of the Nyquist frequency from the CCD images recorded in a 300-kV electron cryomicroscope at a moderately high but practically acceptable microscope magnification.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Bacteriophages / ultrastructure
  • Bacteriorhodopsins / ultrastructure
  • Cryoelectron Microscopy / instrumentation*
  • Cryoelectron Microscopy / standards
  • Image Processing, Computer-Assisted / instrumentation
  • Image Processing, Computer-Assisted / standards

Substances

  • Bacteriorhodopsins