MetaboSign – Technology & Equipment

Agilent Seahorse XF96

The Agilent Seahorse XFe96 analyzer measures in real-time the oxygen consumption rate (OCR) and the proton efflux rate (PER) or extracellular acidification rate (ECAR) of any type of cultured cells. These measurements are performed using sensor cartridges containing oxygen- and proton-sensitive fluorophores, which create a transient microchamber 200 microns above the cell monolayer. Measuring variations in dissolved oxygen concentration and free protons reflects the two primary pathways of cellular metabolism: mitochondrial respiration and cytoplasmic glycolysis.

Additionally, an integrated drug delivery system allows for the sequential and automated addition of up to four compounds into each well at user-defined intervals. This system makes it possible to challenge metabolic pathways, investigate the effects of molecules on metabolism, and test therapeutic compounds.

Using drugs targeting specific components of the respiratory chain, the Seahorse notably enables the measurement of basal and maximal respiration, spare respiratory capacity, and ATP-linked respiration (Mito Stress Test). By using drugs that challenge glycolysis, it also measures basal glycolysis, glycolytic capacity, and glycolytic reserve.

Our Seahorse XFe96 operates with 96-well culture plates, offering immense flexibility for designing various protocols regarding replicates, treatments, choice of injection compounds, and the number of measurements over time.

Potential combinations with other technologies:

• Spectral cytometry: A 35-color panel of immunometabolic markers (NeuroDiderot Cytometry Core Facility) is currently under development.
• Metabolomic NMR: Analysis of metabolites in plasma, culture supernatants or intracellular extracts (Metabo Paris Santé) https://biomedicale.u-paris.fr/rmn-metabo-paris-sante/