The structure of the regular surface layer of Aquaspirillum serpens MW5 has been investigated by electon microscopy supplemented by computer image processing and least-squares analysis. The layer has a ribbed appearance, both on the bacterium and in isolated, negatively stained fragments. However, detailed analysis indicated that the layer was composed of two hexagonal sheets having p6mm symmetry and a = 16 nm. One sheet was staggered by one half repeat along a (1,0) line of the p6nm lattice relative to the second so that, in projection, the pattern of the composite layer was a translational moiré, characterized by a series of ribs spaced 16 nm apart. The ribbed layer had cmm symmetry with a = 32 nm and b = 18.5 nm. Analysis of this pattern indicated that the two p6nm hexagonal sheets were unevenly stained, and this was confirmed by using least-squares methods to simulate the observed pattern by combining two hexagonal patterns. The general structure of the layer was consistent with a role as a selective and protective barrier on the cell surface.