Tissue oxygen plays a crucial role in maintaining tissue viability and in various diseases, including responses to therapy. Useful knowledge has been gained by methods that can give limited snapshots of tissue oxygen (e.g., oxygen electrodes) or evidence of a history of tissue hypoxia (e.g., EF5) or even indirect evidence by monitoring oxygen availability in the circulatory system (e.g., NMR methods). Each of these methods has advantages and significant limitations. EPR oximetry is a technique for direct measurement of tissue pO2, which has several advantages over the other existing methods for applications in which the parameter of interest is the pO2 of tissues, and information is needed over a time course of minutes to hours, and/or for repetitive measurements over days or weeks or years. The aim of this article is to provide an overview of EPR oximetry using particulates to readers who are not familiar with this technique and its potential in vivo and clinical applications. The data presented here are from the experiments currently being carried out in our laboratory. We are confident that in vivo EPR oximetry will play a crucial role in the understanding and clinical management of various pathologies in the years to come.