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This protocol demonstrates the capability of low-field electron paramagnetic resonance (EPR)-based techniques in combination with functional paramagnetic probes to provide quantitative information on the chemical tumor microenvironment (TME), including pO2, pH, redox status, concentrations of interstitial inorganic phosphate (Pi), and intracellular glutathione (GSH). In particular, an application of a recently developed soluble multifunctional trityl probe provides unsurpassed opportunity for in vivo concurrent measurements of pH, pO2 and Pi in Extracellular space (HOPE probe). The measurements of three parameters using a single probe allow for their correlation analyses independent of probe distribution and time of the measurements.

journal of visualized experiments

&lt;em&gt;In Vivo&lt;/em&gt; EPR Assessment of pH, &lt;em&gt;p&lt;/em&gt;O&lt;sub&gt;2&lt;/sub&gt;, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment