The XFe96 analyzer from Seahorse Bioscience measures multiple parameters (O2 and H+) simultaneously using optical biosensors in cell culture microplate assays in real time It is capable of automatically injecting 4 different compounds into the wells, allowing for fast and reproducible measurement of extracellular fluxes before and after injection. Oxygen is consumed during cellular metabolism and extracellular medium is acidified due to excretion of lactic acid produced during glycolysis. O2 and pH flux analysis, combined with injection of compounds targeting metabolism, provides a detailed profile of cellular metabolic phenotype. In the instrument disposable cartridges, embedded with 96 doublets of fluorescent biosensors, are coupled to a fiber-optic waveguide. The waveguide delivers light at different excitation wavelengths and transmits the emitted fluorescent signal through optical filters to highly sensitive photodetectors.
Our Seahorse is housed inside an hypoxic chamber (Custom made by Coy Lab) so that real-time analysis of metabolic function can be performed at a wide range of oxygen concentrations (including hypoxia). This allows for analyzing in vitro cellular and organismal metabolic functions at relevant in vivo oxygen levels.
We equipped our Seahorse with the Agilent XF normalization system providing an even higher level of accuracy and standardization of the cellular assays we perform with the Seahorse. It consists of the Cytation 1 (Biotek, Agilent) and integrated software for automatic imaging and normalization of plates and data.