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Metabolic Analysis of Cells

In 1931 Otto Warburg was awarded the Nobel Prize for his work demonstrating that cancer cells selectively oxidize glucose (glycolysis) rather than energy rich fatty acids compared with  non-cancerous cells. Understanding how the metabolism of cancer cells has changed can be important in their characterization along with the development of new therapies.

The XF Extracellular Flux Analyzer from Seahorse Bioscience is fully-integrated instrument that simultaneously measures the two major energy yielding pathways – aerobic respiration and glycolysis – in a convenient, microplate format. The system is ideal for measuring metabolic changes in cells in response to ligand stimulation or drug treatment.

Using disposable fluorescent optical sensors and multiplexed fiber optics, the instrument offers:

  • Simultaneously measure oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) in as little as five minutes
  • Perform time-resolved measurements of a single population of cells over a period of minutes, hours or even days
  • Re-use cell samples after completing an XF measurement

The PCL now offers access to the XF Analyzer as a walk-up machine or as a complete service. Training, protocols and disposables (plates and sensors) are available through the PCL.  Please contact Dr. Andrew Stephen (301-846-1634) for more information.

One of the standard assays that is performed in the training using the XF Analyzer is the Mitochondrial Stress Test.  Inhibition of specific electron carrier complexes in the mitochondria can provide insightful information on basal respiration, ATP turnover, proton leak, and maximal respiration.

  • Oligomycin inhibits Complex V
  • FCCP decouples the proton gradient
  • Rotenone/Antimycin A inhibits Complex III
Cells are cultured and the specific inhibitors are injected at defined times. The OCR and ECAR are measured in response to inhibitor treatment. The data shown below is a mitochondrial stress test performed by PCL on a pancreatic cell line showing the effect on OCR of the inhibitors.

 

 

Page last updated August 8, 2012 @ 4:27 pm