JoVE Encyclopedia of Experiments
Cancer Research
0 views • 4:39 min • April 30th, 2023
- Extracellular flux assay helps study cell metabolism by measuring the two major energy pathways-- glycolysis and mitochondrial respiration. During glycolysis, cells metabolize glucose and secrete lactate into the extracellular space along with protons, determined by measuring the Extracellular Acidification Rate or ECAR. During mitochondrial respiration, cells utilize oxygen for ATP production, measured by determining the Oxygen Consumption Rate or OCR.
Cancer cells exhibit high glucose uptake as they alter their metabolic rate to adapt and maintain unregulated cellular proliferation. To analyze metabolism in cancer cells, first assemble a flux analysis microplate containing adherent cancer cell monolayer and a hydrated sensor cartridge. Transfer the plate assembly into a flux analyzer. Within the analyzer, the sensor probes descend to create a transient microchamber. To measure extracellular flux, the injection port introduces test compounds into the microplate at preset time points.
Based on cell metabolism, the concentrations of oxygen available and protons released in the microchamber fluctuates. Two fluorophores embedded at the base of the cartridge simultaneously senses and
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