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Seahorse XF Pro Analyzer

Overview:

XF_ProThe Seahorse XF Pro Extracellular Flux Analyzer allows for real-time quantitation of two major energy-yielding pathways in the cell: mitochondrial respiration and glycolysis. This instrument offers considerable improvements compared to former methodologies. XF Pro analyzers do not require radio-labeled substrates, and unlike the classic Clark electrode, it is possible to make high-throughput measurements of cellular/mitochondrial oxygen consumption on 90 samples simultaneously in a microplate format.

The XF Pro uses disposable fluorescent-based optical sensors to measure oxygen consumption rate (OCR), extracellular acidification rate (ECAR), and proton efflux rate (PER) directly in the media of microplates. OCR and PER reflect the flux through mitochondrial oxidative metabolism and glucose oxidation, respectively. Changes in cellular/mitochondrial metabolism in response to specific inhibitors or other substances can also be determined. Built-in injection ports surrounding each of the wells in a microplate allow sequential delivery of up to four compounds (e.g., fatty acids, glucose, pyruvate, succinate, FCCP, oligomycin, rotenone, antimycin) to samples during the course of an experiment.

An XF Spheroid Microplate is available to assess functional metabolism in 3 dimensions on spheroids (1-100 spheroids per well, for glucose or oligomycin response studies). 

The XF Technology:

  • Measures oxygen consumption rate (OCR), extracellular acidification rate (ECAR) and proton efflux rate (PER) of living or frozen samples simultaneously in real time.
  • Samples assayed on the XF analyzer: Cell lines, primary cells, isolated mitochondria, and 3D spheroids.
  • Typically requires only 5,000 to 40,000 cells per well in 96-well plates.
  • Up to four test compounds (drugs or substrates) may be added automatically to each well. Measurements are performed before and after each compound is added.
  • Measurements may be repeated multiple times to measure kinetic responses.
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Department of
Pharmacology, Physiology, & Neurobiology
College of Medicine
231 Albert Sabin Way
Cincinnati, OH 45267-0575