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For more information read the application note Seahorse XF Metabolic Pathway Determination Assay.
XF Bioenergetic Applications Provide a New Window on Cell Metabolism
Seahorse XF instruments measure energy utilization in living cells, simultaneously quantifying mitochondrial respiration and glycolysis in real time. The technology offers a robust and simple method for studying substrate utilization, mitochondrial function, energy expenditure and cell quality in microplates, without the use of large number of cells, flasks, electrodes, dyes, radioactive materials or lysis of cells that is typical of other methods.
Substrate Utilization
Feeding cells substrates such as glucose, pyruvate, galactose, palmitate and oleate cause fluctuations in metabolic pathways. XF measurements of cells exposed to various substrates provide estimates of metabolic fluxes with more information than those obtained using radiometric methods. The advantages of XF assays are higher throughput, reduced sample sizes and substantially-improved kinetic resolution of both energy producing pathways with rates that are typically obtained within several minutes.
Mitochondrial Function
Historically, isolated mitochondria were used to study bioenergetics, drug toxicity and diseases including obesity, diabetes, neurodegeneration and cardiomyopathy because alternative cell-based methods were not available. With the introduction of XF assays, investigators can measure mitochondrial function in living cells which offer a better correlation to "in vivo" function. Primary, adherent and suspension cells are routinely used.
Energy Expenditure
In response to a changing extracellular environment, disease or drug exposure, cells remodel bioenergetic pathways in a matter of minutes as they strive to maintain appropriate levels of ATP necessary for their biological function. By measuring the two major energy producing pathways of the cell, mitochondrial respiration and glycolysis, XF assays provide a more comprehensive picture of the kinetics of metabolism that drives cellular function.
Cell Quality Assessment
Simple viability tests do not provide detailed information about the functional potential or "vitality" of cells; and the changes they detect are hours behind the actual energetic changes in the cell. With XF assays, the bioenergetic state of a cell population can be easily and non-invasively measured. The resulting bioenergetic data is indicative of the quality and functional potential of the cells. Pre-testing cells prior to running critical assays can save money, compounds, reagents and time.
Cell Line Reference Chart
Here is a reference chart of cells types that have been successfully analyzed in XF experiments. Data is from our customers and this chart will be updated as more information on cell types is received. Potentially any cell type can be measured in an XF assay; however, because of intrinsic differences between cells derived from different tissues distinct protocols are necessary to measure them under appropriate conditions.