Multicolor FISH using whole-chromosome painting and FISH banding with partial chromosome painting probes are, for technical reasons, not suited to the characterization of small centromeric and pericentromeric rearrangements. Thus, probe sets for characterizing the different human centromeres, and others that enable studies of the near-centromere regions of human chromosomes, were established. Such centromere-specific multicolor (cenM-) and subcentromere-specific (subcenM-) FISH approaches are presented in this chapter. The preparation and use of these probe sets is reported.