Correlation averaging and structural changes in GP32*I crystal unit cells

Abstract

We compared correlation and Fourier averaging by analyzing 400 kV spot-scan electron images of iceembedded gp32*I crystals from a previous study using correlation averaging and multivariate statistical analysis (MSA). The set consisted of 9 spot-scan electron images (Fig. 1) from different micrographs. Unit cells of gp32*I crystal in each spot-scan image were located using cross-correlation map ofthe whole image with its central portion (an area of up to 10 unit cells). Each correlation peak in the map served as a center for picking up a patch of the same size as the reference used for mapcalculation. After averaging of the patches a new reference with improved signal-to-noise ratio was obtained and procedure was iterated several times using progressively smaller reference sizes. Final reference size was about 1-2 unit cells. This permitted us to follow local lattice variations. The technique does not use any assumption of crystallinity of the specimen and can be applied for highly distorted crystal lattices. Each spot-scan image was processed independently of the others. After the iterativeprocedure a final set of averaged images was obtained and structure factor amplitudes and phases were calculated from them using standard crystallographic procedures followed by merging all the spectra. Using the set of structure factors a projection map was generated which looked very similar to the 4 Å resolution map obtained using Fourier averaging technique (Fig. 2).