MCW Cancer Center Bioenergetics Shared Resource

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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 theory behind this instrument utilizes 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”.

  • The cost per experimental plate is $125.00.
  • This price includes the compounds utilized in a basic mitochondrial stress test (oligomycin, FCCP, and ANT.A/Rotenone)

 

 

Certain stressors can have different effects on oxygen consumption in various cell lines. Examples of oxidative stressor can be seen below:

 

                                         Graphical Abstract from Dranka et. al FRBM (2011, in press)

 

 

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

 

If you would like to schedule a time to use the XF-96, please call Steve Komas at 414-955-4059, or email him at skomas@mcw.edu.  Also, please click the link below to find out if the XF-96 is available to use at your desired time.

 

XF-96 Google Calendar

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Page Updated 03/27/2014