CD45 is a glycoprotein expressed on all lymphohematopoietic cells. Its expression increases during normal B‐cell differentiation and remains stable on mature cells. Although it is widely known that CD45 antigen expression is decreased in B‐acute lymphocytic leukemia (ALL), only scarce and contradictory information is available on CD45 expression on mature B‐cell malignancies. In healthy adults (n = 15), CD45 expression on B lymphocytes was lower than that on T cells. In patients with chronic lymphocytic leukemia (CLL; n = 22), CD45 expression on malignant cells was lower than that on the whole lymphocyte population of healthy adults (n = 28) and on normal B lymphocytes (n = 15). In 6 of the 22 CLL patients, the malignant cell population could be separated from the normal lymphocyte population on the CD45‐side scatter (SSC) plot. In 16 CLL patients, there was some degree of overlap between the malignant and normal cells with respect to CD45 expression. For these patients, there was an inverse correlation between CD45 expression on the whole lymphocyte population and the percentage of malignant cells in this population. In two patients with mantle cell lymphoma (MCL), CD45 expression on the malignant cells appeared lower than that on normal B cells and on the whole lymphocyte population. In six patients with hairy cell leukemia (HCL), CD45 expression on hairy cells was comparable to that on the whole lymphocyte population of healthy adults, but slightly higher than that of normal B cells. Evaluation of CD45 expression may help to characterize mature B‐cell malignancies. Cytometry (Comm. Clin. Cytometry) 46:336–339, 2001. © 2001 Wiley‐Liss, Inc.