Determine extracellular acidification and oxygen consumption in real-time
The metabolic condition of a cell sheds light on its state of health. The rate of glycolysis is one metabolic factor impacting cellular physiology and pathology. For instance, cancer cells exhibit a higher rate of glycolysis compared to healthy counterparts. Since lactate is produced in the course of glycolysis and subsequently released into the extra-cellular space, it acidifies the surrounding medium. The so-called extracellular acidification is hence used to monitor glycolytic flux.
A second metabolic parameter is the consumption of molecular oxygen (O2) as it reflects oxidative phosphorylation. In cancer cells this pathway of energy production is decreased. Furthermore changes in oxygen consumption may indicate mitotoxicity of drugs.
Agilent Technologies provides probes that are sensitive to either pH or O2 and reliably trace both metabolic processes: glycolysis and respiration. Both probes can be used simultaneously, assuring maximal information output from your experiment.
The CLARIOstar microplate reader is the ideal tool to measure these metabolic assays as it easily detects the probes and can be equipped with an Atmospheric Control Unit (ACU). This accessory allows working at desired oxygen and carbon dioxide concentrations that resemble physiological conditions. It further enables to work at steady O2 and CO2 concentrations, a requirement for determination of extracellular acidification (ECA) and oxygen consumption rate (OCR).
Fig. 1: Monitoring O2 concentrations in samples containing 3D Hep2 cells in response to decreasing atmospheric O2 conditions realized by the CLARIOstar ACU.
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Real-time measurement of intracellular O2 in mammalian cells
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