Biological membranes comprise thousands of different lipids, differing in their alkyl chains, headgroups, and degree of saturation. It is estimated that 5% of the genes in the human genome are responsible for regulating the lipid composition of cell membranes. Conceivably, the functional explanation for this diversity is found, at least in part, in the propensity of lipids to segregate into distinct domains, which are important for cell function. X-ray diffraction has been used increasingly to characterize the packing and phase behavior of lipids in membranes. Crystalline domains have been studied in synthetic membranes using wide- and small-angle X-ray scattering, and grazing incidence X-ray diffraction. Herein we summarize recent results obtained using the various X-ray methods, discuss the correlation between crystalline domains and liquid ordered domains studied with other techniques, and the relevance of crystalline domains to functional lipid domains in biological membranes.