The presence of stem-like cells in tumors reflects the invasive character of the disease; however, their identification is controversial. We investigated the distribution of CD133, CD44 and CD24 using histological sections and tissue microarrays (TMAs) of human colon adenocarcinoma obtained from patients with and without lymph node metastases and/or liver metastases. Immunohistochemical staining was combined with nuclear staining and evaluated quantitatively using image analysis software. Sections of normal colon mucosa, the primary tumor, lymph node, and liver also were analyzed qualitatively and compared to the quantitative method, which was more accurate. In most tissues, the expression of CD44 and CD24 was relatively low compared to CD133, with some variations. CD133 also was expressed in the normal colon mucosa and to a lesser degree in normal hepatic parenchyma. Liver metastases exhibited significantly greater CD133 staining compared to normal colon mucosa, primary tumor and lymph node metastases. Moreover, lymph node metastases obtained from patients with liver metastases expressed significantly greater CD133 staining than those obtained from patients without liver metastasis. Our data suggest that CD133 expression in lymph node metastases may be of value for prognosis of the disease.