Real-Time Intracellular Oxygen Assay

AMSBIO has introduced MitoXpress Intra, a novel intracellular oxygen assay based on a 96 or 384-well fluorescence plate reader-based approach that facilitates the real-time assessment of transient changes in cell respiration, oxygen gradients and physiological responses across a range of cell models.

Molecular oxygen is the key substrate of aerobic metabolism. Knowledge of cell oxygenation is therefore central to a detailed understanding of the cellular metabolic response to a particular treatment or manipulation. Traditionally intracellular measurements have proven very difficult, requiring the use of invasive, laborious, low-throughput, technically challenging techniques which in turn have limited the use of such measurements within biological research. Using a MitoXpress Intra probe researchers can, for the first time, monitor in real time molecular oxygen within the cell monolayer on a plate reader in a non-invasive, high-throughput manner.

MitoXpress Intra is a nanoparticulate metalloporphryin-based, phosphorescent, oxygen sensitive probe that is used to monitor intracellular oxygen in populations of live mammalian cells by plate-based, time-resolved fluorometry. The assay is based on the ability of molecular oxygen to quench the excited state of the probe. The probe is taken up via non-specific energy dependent endocytosis and, after washing, the cells are monitored on a time-resolved fluorescence plate reader. Probe phosphorescence is quenched by intracellular oxygen in a non-chemical, reversible manner allowing the measurement of average intracellular oxygen levels and facilitating real-time monitoring of relative changes in cellular oxygen consumption. Probe signal increases with a reduction in intracellular oxygen and decreases with an increase in intracellular oxygen.

The MitoXpress Intra assay is optimized for use on standard 96 / 384-well plates and time resolved fluorescence plate readers enabling high throughput analysis of multiple samples in parallel. Data produced from the chemically stable, inert MitoXpress Intra probe is complementary to other intracellular parameters, such as ROS and mitochondrial membrane potential.

MitoXpress Intra is a powerful new tool that has been shown to facilitate the real-time assessment of transient changes in cell respiration, oxygen gradients and physiological responses across a range of cell models. Specifically, it facilitates the measurement of cellular oxygenation; a critical parameter across many fields of research including hypoxia and cancer metabolism. For more information,visit www.amsbio.com/metabolism-oxygen-consumption-and-glycolysis-assays.aspx.