The use of orthogonal projection to visualize mosaic domains from topographic data collected on protein crystals

Owing to the relatively low intensity of diffraction and resulting low contrast topographic images, and also the richly detailed domain structure, it can be difficult to extract individual mosaic domains (size and shape) for protein crystals. Here, orthogonal projection was tested and found to be a superior approach over previous methods to extract mosaic domain information from a fine‐sliced topographic sequence of images. The topographic sequence was collected at room temperature from a lysozyme crystal with a low mosaicity. Orthogonal projection can be applied to image sequences that are spatially invariant and composed of linearly additive image formation processes. The domains were determined by using a particle swarm optimizer to fit Gaussians to the integrated intensity profile of the reflection as a function of angle, although any basis function could have been used. This optimization method was more amenable to automation and converged to the global minimum more efficiently. The number of domains can normally be determined by visually inspecting the integrated intensity profile; this only needs to be done once for each crystal, because the number of domains remains constant during collection, and not for each reflection. The results can be used to provide a picture of the internal structure of the crystal and may be useful to aid in the improvement of crystal growth techniques.