A growing appreciation for the importance of hypoxia in tumor progression and response to treatment has driven efforts to develop methods that could be used routinely in the clinic to identify tumors containing hypoxic cells. The ideal method would be noninvasive and could be used both before treatment to determine the presence of hypoxia and during therapy to assess tumor reoxygenation. Although this goal is being approached, there are still questions about how best to measure tumor oxygenation and whether noninvasive imaging methods can provide the necessary sensitivity. Analysis of hypoxia at the level of the individual cell can provide the following information that cannot be obtained in other ways: the degree of hypoxia, the lifetime of hypoxic cells, and the dynamic nature of hypoxia. This review will describe methods that have been used to detect hypoxia in individual cells, the relation between these measurements and patient response to treatment, and indicate where these methods can provide important additional insights into the consequences of tumor hypoxia.