Efficient detection of somatic mutations is important for the development of clinical molecular diagnostic assays. However, the detection of somatic mutations in tissue is confounded by dilution of the tumor cell population by normal cells. Laser microdissection allows enrichment for tumor-associated genetic alterations to take place at the level of cell selection, eliminating the need to enrich for mutant alleles after amplification. In this chapter a method is described for somatic mutation analysis using cells acquired by laser capture microdissection.