Tumor oxygenation determines the efficacy of radiotherapy, but there is no non-invasive way to image this parameter. Since gradient recalled echo (GRE) images are sensitive to blood deoxyhaemoglobin concentration ([dHb]) they could have a role in assessing tumor oxygenation. In brain, linear relationships have been demonstrated between brain tissue R2* relaxation rate and tissue [dHb] or oxygen saturation, but in tumors, vascular and tissue heterogeneity, and the presence of simultaneous oxidative and glycolytic metabolism, complicate the analysis. We have studied the effects of vascular challenge in a rat prolactinoma tumor model by MR imaging and spectroscopy and comment on the implications of these results for calibrating GRE images for blood or tissue pO2.