How Agilent Seahorse XF Analyzers Work (2024)

Once a measurement is completed, the probes lift which allows the larger medium volume above to mix with the medium in the transient microchamber, restoring cell values to baseline. An integrated drug delivery system allows sequential addition of up to four compounds per well at user-defined intervals.

Because Seahorse XF measurements are non-destructive, the metabolic rate of the same cell population can be measured repeatedly

over time while up to four different drugs are injected sequentially into each well. Total assay time is typically 60 to 90 minutes. Upon completion of an XF assay, other types of biological assays such as cell viability can be performed on the same plate. To maintain normal cell physiology, a temperature control system maintains the cellular environment at 37°C (or other pre-set temperature).

Seahorse consumables are developed specifically for use with the Seahorse XF Analyzer. They provide sensitive, kinetic measurements to simultaneously analyze the two main metabolic energy pathways: mitochondrial respiration and glycolysis.

Seahorse XF FluxPaks include sensor cartridges, tissue culture plates, calibrant, and utility plates for performing calibration. The sensor cartridges contain solid state sensor probes for detecting oxygen and proton levels in cell culture media. They also have integrated injection ports for introduction of stimuli, inhibitors, and substrates during the assay which enables monitoring transient cellular responses in real time. The XF tissue culture plates are provided in 8-well, 24-well and 96-well formats. Specialized cell culture plates have been designed for spheroids and islets. Every lot of XF Fluxpaks and culture plates is 100% quality controlled for performance and lot-to-lot consistency.

Seahorse XF kits, reagents and media are designed for performing a wide range of assays with XF analyzers and XF Fluxpaks, to study a range of cellular function, including metabolic phenotyping, ATP production, mitochondrial function, fatty acid oxidation, and glycolysis in real time. They are standardized, optimized open-and-go products that allow to achieve consistent results, reduce assay complexity and improve research productivity. Each lot of products are rigorously tested and validated with cells to deliver high performance and low lot-to-lot variation. Together with Agilent's excellent technical support, these kits, reagents and media will greatly enhance your overall experience of performing XF assays.