Since individual chromosomes can be accurately identified by new banding techniques, atebrin fluorescence was used for chromosome analysis in six cell lines and two primary outgrowths derived from human malignant melanoma. Gross aneuploidy was seen in all specimens, but each culture contained at least 1 distinctive marker chromosome specific for that cell line in 87 to 100% of metaphases. One of the primary explants contained a marker that was demonstrable in fresh tissue and persisted through 2 weeks of culture. The same marker was found in all metaphases from 2 different metastases, but skin fibroblasts from the same patient had a normal chromosome complement. No common marker for human melanoma was found, but in 6 of the 8 cultures the most frequently found marker was formed by a brightly banded chromatid addition. Relative polysomy for Chromosome 7 was found in 7 of the 8 cultures and, for Chromosome 22, in 8 of the 8 cultures. The frequency of polysomy of Chromosomes 7 and 22 was significant at the 5% level.