High-throughput real-time analysis of cell oxygenation using intracellular oxygen-sensitive probes

Knowledge of in situ oxygenation of cells in 2D and 3D cultures offers important insights into the impact of oxygen on cellular function. Here we outline how such measurements can be performed in 2D cultures of adherent cells and also in cells cultured on 3D scaffolds. Measurements are performed on conventional time-resolved fluorescence plate readers using the intracellular oxygen-sensitive probe MitoXpress(®)-Intra. We also illustrate how the impact of drug treatment on cell oxygenation can be assessed and how the link between oxygenation and glycolytic metabolism can be examined.