DNA synthesis of mouse L cells in vitro was studied by immunofluorescence and immunoperoxidase techniques using anti-BUdR antibody to identify cells which incorporate BUdR. The fraction of fluorescence- or peroxidase-positive cells agreed well with ^3H-thymidine labeling index. In synchronized cell populations, DNA synthesis rate as measured by fluorescence intensity increased from immediately after block and after plateau of a 4-hr period returned to the initial level in 8 hr. At the mid S phase when DNA synthesis rapidly increased, fluorescence intensity of individual cells varied significantly.