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Fluxomics

As part of a joint initiative between RESTORE, CRCT and the MetaToul platform, CERT houses an in vivo fluxomics facility that includes:
- two modules allowing the realization of isotopic tracing experiments (stable isotopes: 13C, 15N, etc) in vivo or in vitro
- the equipment necessary for the collection of samples and the extraction of metabolites
- a culture chamber under controlled oxygen pressure to understand the effect of oxygen partial pressure variations on metabolism
- a respirometry tray to measure oxygen consumption flows
Metabolome, lipidome and isotope labeling analyses are performed on the MetaToul platform, which has an instrumental park (mass spectrometry, NMR) and unique expertise in this field in France. The whole set-up and the associated expertise represent also a unique configuration in France for in-depth studies of metabolic dynamics at different scales (from the cell to the whole organism).

in vivo Fluxomics module
It contains the infrastructure and equipment necessary to perform a complete in vivo isotope tracing study on mice:
- Establishment using animals for scientific purposes approved by the DDPP
- Gas anaesthesia tables for the catheterization of mice
- Device for perfusion of 13C-labeled substrates (or other stable isotopes) on vigorous mice (syringe pumps, portals, swivels)
- Tissue collection and preparation, sample preparation for analysis.

in vitro Fluxomic module
This module has the necessary equipment to carry out an in vitro isotope tracing study in its entirety: cell culture under controlled oxygen atmosphere (see culture under controlled oxygen atmosphere), labelling experiments with 13C-labelled metabolites (or other stable isotopes), sampling and extraction of internal (endo-metabolome) or external (exo-metabolome) metabolites:
- Fully oxygen controlled atmosphere chamber (X-Vivo, Biospherix)
- Various equipment for endo- and exo-metabolome extraction
- Quenching system (immediate blocking) of the in vitro metabolism

Oxygraphs
The Oroboros type oxygraphs present on the CERT platform are very sensitive instruments due to their hermetic tanks and Clark probes. They allow the study of mitochondrial activity on tissues (homogenates, permeabilized), cells and isolated mitochondria using different substrates and drugs. The Oroboros type oxygenographs are not limited in the number of injections per experiment which allows a more precise study of mitochondrial activity:
2 Oroboros oxygraphs
2 independent tanks per oxygenograph
Métabolisme, adipocytes, métabolomique, fluxomique, traçage isotopique in vitro/in vivo, culture cellulaire en hypoxie, activité mitochondriale

- provides advice in the development of experiments,
- trains the users,
- assists users in the analysis and interpretation of their results,
- assists users in the implementation of research and development projects, in the form of collaboration.