Patch averaging of electron images of GP3*I crystals with variable thickness

The combination of Fourier and correlation averaging techniques with multivariate statistical analysis and classification, a method known as patch averaging, is used to analyze untilted and tilted images of negatively stained GP32*I crystals, which exhibit variable thicknesses in a single crystal. Within a single image, coherent areas of the same apparent thickness can be distinguished from areas of differing thicknesses. Analysis using the phase relationships among symmetry-related reflections from reconstituted images obtained from untilted micrographs confirms the ability of the method to classify these variable thicknesses properly. Furthermore, the phases from some of the reconstituted images obtained from both untilted and tilted micrographs were found to match well with the phases in a previously determined three-dimensional data set of this crystal with pg symmetry along the crystallographic b axis. These results indicate the utility of the patch averaging procedures in the structural determination of protein crystals with different thicknesses.