The present study investigates the correlation between tumor oxygen availability and radiosensitivity following oxygen manipulation. Previous work has shown that tumors may contain both diffusion- and perfusion-limited hypoxic cells. Recently, the combination of nicotinamide (NIC) administration plus carbogen breathing has been proposed as a means of targeting both hypoxic cell subpopulations. Intravascular HbO2 saturations were measured for KHT murine sarcomas following either NIC, carbogen breathing, or the combination, and compared with determinations of tumor cell survival under matched conditions. The percentage of vessels > or = 25% HbO2 increased significantly for both the carbogen and NIC-carbogen combination, while remaining unchanged from controls following NIC. These findings contrast with the survival data, where all treatments showed identical cell survival. A possible explanation is that different proportions of clonogenic versus nonclonogenic cells may be oxygenated by the alternative treatments. Thus direct determinations of alterations in tumor oxygenation may not reflect corresponding changes in radiosensitivity.