Three-dimensional reconstruction of imperfect two-dimensional crystals

An outline is given of the general methodology of 3D reconstruction on the basis of correlation averages of the 2D projections: this hybrid real space/Fourier space approach substantially alleviates one of the most serious limitations on obtaining high resolution 3D structures, namely crystal distortions. The paper discusses some of the technical problems involved, namely optimisation of tilt increments, a posteriori tilt angle determination, extraction of lattice line data from averaged unit cells, and stain/protein boundary determination. The approach is illustrated by application to a 2D crystal from a bacterial cell envelope.