Pimonidazole, a "hypoxia marker" normally delivered i.v. or i.p., was instead administered in the drinking water of tumor-bearing mice. As pimonidazole exposure was increased from 3-96 h ad libitum, both the fraction of hypoxic tumor cells and the relative number of pimonidazole adducts in those cells increased. Furthermore, the sustained ingestion of pimonidazole revealed a larger hypoxic fraction than did a single injection of an alternative hypoxia marker, CCI-103F. The "additional" hypoxia seen with longer-term oral administration apparently reflects the inclusion of transiently hypoxic tumor cells. Thus, in addition to its convenience and versatility when compared with hypoxia marker injection, oral administration of pimonidazole appears to permit identification of all of the physiologically and therapeutically relevant hypoxic tumor cells.