An assay for local quality in cryo-electron micrographs of single particles.

High quality of the cryo-electron micrographs is of crucial importance for the success of single particle three-dimensional reconstruction methods. In analyzing some micrographs from cryo-electron microscopy specimens, we found an extraordinary variability, within the same micrograph, in the appearance of particles. We developed a method for analyzing the variability of local image quality, using correspondence analysis of local power spectra. With this technique, we discovered a strong systematic variation of the envelope modulating an otherwise unchanged contrast transfer function. The underlying causes may be uncontrollable effects, such as variations in the thickness of ice, instability of the holey carbon, and charging. The method of assaying, resulting in "local quality maps", may be useful as a general tool for screening micrographs used as input for reconstructions.