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Redox & Bioenergetics Shared Resource and Free Radical Research Center

Seahorse Bioscience Extracellular Flux Analyzer

The Seahorse XF-96 measures the rate of change of analytes dissolved oxygen and pH in media immediately surrounding living cells cultured in a microplate. Changes in the extracellular media are caused by the consumption or production of analytes by the cells. Therefore, a sensitive measurement of the media flux can be used to determine rates of cellular metabolism with great precision and in a totally noninvasive, label free manner. The analyzer can make measurements in as little as five minutes. Cells can be profiled over a period of minutes, hours, or days.

The cost per experimental plate is $200. This price includes the compounds utilized in a basic mitochondrial stress test (oligomycin, FCCP, and ANT.A/Rotenone).

Contact us to schedule time to use the XF-96.

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xf-96analyzer

The theory behind this instrument is based on the fact that normal cells rely on mitochondrial oxidative phosphorylation (OXPHOS) to produce ATP (34 molecules of ATP/one molecule of glucose) as a source of cellular energy. On the other hand, most cancer cells utilize aerobic glycolysis which is energetically inefficient, to generate ATP (2 ATP/one molecule of glucose) with a decreased use of TCA cycle. Subsequently, tumor cells are glycolytic and actively take up much more glucose. Glucose is converted to G-6-P by high intracellular levels of hexokinases. This metabolic shift from OXPHOS to aerobic glycolysis with increased lactate production is known as the "Warburg effect."

oxygenconsumptionpicture

Certain stressors can have different effects on oxygen consumption in various cell lines. Examples of oxidative stressors are shown in this figure (Dranka et al. FRBM 2011, 51(9):1621-35).
ocrandecarexplanation

The Seahorse XF-96 has the ability to analyze both OCR and ECAR simultaneously without disrupting the cells.