Mitochondrial Function using the Seahorse Bioscience Extracellular Flux (XF) Analyzer

The XF Analyzer measures oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) at intervals of approximately two - five minutes. OCR is an indicator of mitochondrial respiration, and ECAR is predominately the result of glycolysis.

Real-time measurements of OCR and ECAR are made by isolating an extremely small volume (less than 7µl) of medium above a monolayer of cells within a microplate.

Cellular oxygen consumption (respiration) and proton excretion (glycolysis) causes rapid, easily measurable changes to the concentrations of dissolved oxygen and free protons in this transient microchamber, which are measured every few seconds by solid state sensor probes residing 200 microns above the cell monolayer.

The instrument continues to measure the concentrations until the rate of change is linear and then calculates the slope to determine OCR and ECAR, respectively.

The metabolic phenotyping core has the capability to assess oxygen consumption and CO2 production for measures of state 3 (ADP- stimulated), state 4 respiration, and uncoupled (FCCP- stimulated) respiration in both cultured cells, mitochondria isolated from various tissues, and isolated skeletal muscle fibers.

In addition, the core has the ability to assess ATP synthesis and reactive oxygen species production in isolated mitochondria.

Virginia Tech Metabolic Phenotyping Core
Integrated Life Sciences Building
1981 Kraft Drive
Blacksburg, Virginia 24060