In the majority of in vitro studies cells are cultured and studied at ambient oxygen - ignoring the oxygen gradient between the atmosphere and the medium as well as the gradient between the medium and the intracellular cell environment. As the level of available oxygen significantly influences cell physiology any data obtained from cell-based in vitro studies should be related to the level of cellular oxygenation.
To that end Agilent Technologies has developed the MitoXpress Intra - Intracellular Oxygen assay kit. The assay uses an oxygen-sensitive fluorescent probe. Oxygen quenches the phosphorescent emission from the probe, such that the measurement signal is proportional to the oxygen concentration.
In this application note we show a MitoXpress Intra assay is performed on the CLARIOstar® microplate reader. The reader is equipped with an atmospheric control unit (ACU) in order to reproduce either optimal physiological conditions or hypoxic conditions for 2D and 3D cell-based assays.