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COMPÉTENCES

 

 

Modèles

Principaux modèles utilisés actuellement au LBFA :

    • Hépatocytes isolés incubés ou périfusés
    • Cardiomyocytes isolés
    • Cœur isolé perfusé
    • Cultures primaires et lignées
   
• Exposition à l'hypoxie, à l'hyperoxie
    • Exercice, entraînement
    • Régimes enrichis ou restriction calorique

 

 

Techniques

Principales techniques maîtrisées au LBFA :

    • Mesure du statut énergétique des cellules
    • Oxygraphie sur mitochondries isolées ou cellules perméabilisées
    • Mesure de production de ROS mitochondriaux
    • Histochimie
   
• Enzymologie
    • Electrophorèse de protéines et d'acides nucléiques
    • Expression génique et protéique (western blot, PCR en temps réel)
    • Clamp euglycémique hyperinsulinémique
    • Dosages de nucléotides par HPLC
    • Tests comet
   

 

Infrastructure. All in-house material resources and technical facilities is shared by the teams and can be used without internal billing. This is maintained by an overhead of 25% on all obtained grants (apart of salaries and investments). During the current contract, the technical equipment has been structured into several platforms. These will be combined into seven facilities and centralized with the launch of the new BEeSy research building, where proteomics, cell culture and animal facility will obtain dedicated space. Each facility is managed by 1-2 permanent LBFA members.

  • Proteomics I - recombinant protein expression and purification, interactomics (yeast-two-hybrid, surface plasmon resonance); (phospho)-proteomics (2D-PAGE, DIGE) developed during the current contact.
  • Proteomics II -mass spectrometry - the Promethée facility joining LBFA in the next contracting period (physically remaining at the CHU campus): mass spectrometry, MALDI and electrospray MS, identification of proteins & protein secondary modifications.
  • Cell culture - primary cells & cell lines as model systems.
  • Animal facility - rodent models, animal experimentation (exercise, nutrition, toxicology - extended in the current contract); will be complemented by an insect (mosquito) facility (for LECA); part of the Grenoble IBISA network of animal facilities.  
  • Imaging & cytometry - fluorescence & confocal microscopy, cell sorter/FACS, development of intracellular fluorescent sensors and methodology for bioenergetic applications; joined the Grenoble IBISA network of cellular imaging facilities.
  • Metabolic analysis - numerous applications that are a specialty of LBFA: respirometry, cell perifusion, perfused organ (heart, liver), multiscale hypoxia (protein, cell, animal model), nitro-oxidative  stress markers, enzyme activity, etc.; development of (phospho)metabolomics envisaged in the next contracting period
  • General services - Stocks (chemistry, labware, samples; cold/frozen stocks), chemistry handling (scales, pH meters, media preparation), centrifuges, central washing plant; also spectrometers and PCR.