XFe-24 Extracellular Flux Analyzer (Seahorse Bioscience)

The XFe-24 flux analyzer measures real time kinetic of the media flux in live cellular environment to determine in vitro oxygen consumption rate (OCR), and extracellular acidification rate (ECAR), in order to assess cellular metabolic functions such as oxidative phosphorylation, glycolysis, and fatty acid oxidation. XFe-24 assays apply to all fields of biomedical research. The plate accommodates 24 individual wells, and up to 4 different test compounds (for example, Oligomycin, 2-DG, FCCP, Rotenone) can be injected separately into each well of real time kinetic experiment.

A short video introduction can be found here .

Materials required and recommended for a successful XFe24 assay.

  • Agilent Seahorse XFe24 Analyzer with Controller
  • Non-CO2 Incubator (37°C)
  • Agilent Seahorse XFe24 FluxPak (Part # 102340-001) or Agilent Seahorse XFe24 FluxPak mini (Part # 102342-100)
  • Calibrated pH Meter
  • Inverted Phase Contrast Microscope
  • P20 and P200 Pipettes
  • P20 and P1000 Pipette tips
  • Appropriate XF Assay Media (for guidance in choosing and preparing the proper assay media, see: media selection guide and Preparation of XF assay media) .

Science behind the assay

  1. Seeding Adherent Cells in Agilent Seahorse XF24 Cell Culture Microplates.
  2. Using the Hydro Booster to Improve Hydration and Data Quality (This procedure must be performed 1 day prior to using the Sensor Cartridge for a XF Assay).
  3. Preparation of XF assay media.
  4. Washing Adherent Cells in Agilent Seahorse XF24 Cell Culture Microplates
  5. Loading the Agilent Seahorse XFe24/XF24 Sensor Cartridge with compounds.
  6. Cell Characterization using the Seahorse XF Cell Energy Phenotype Test



Cancer Biology
Other pathways involve changes of glycolysis and mitochondria respiration

Typical data