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.
Key wordsHypoxia Mitochondrial function Cell metabolism Oxidative phosphorylation Time-resolved fluorescence Intracellular oxygen Oxygen-sensitive probe Oxygenation Electron transport chain High throughput
- 10.Valeur B (2002) Molecular fluorescence: principles and applications. Wiley-VCH, New York, NYGoogle Scholar
- 14.Hynes J, Natoli E, Will Y (2009) Fluorescent pH and oxygen probes of the assessment of mitochondrial toxicity in isolated mitochondria and whole cells. In: Maines M, Costa LG, Hodson E, Reed JC (eds) Current protocols in toxicology. Wiley, New York, NY, pp 40:42.16.41–42.16.22Google Scholar