Intravital Two Photon Microscopy

Team

  • Dr Franck DEBARBIEUX, Ph.D. (MCU AMU)
  • Dr Ivo VANZETTA, Ph.D. (CR, CNRS)
  • Pr Geneviève ROUGON, Ph.D. (DRCE émérite CNRS)

Situation

  • Labs : ground and first floor
  • Offices : 1st floor

Contact

Purpose & mission

The Optical Department aims to provide a unique characterization of neuroinflammatory processes at play in mice models of neuropathologies (glioblastoma brain tumors, multiple sclerosis, CNS trauma).

This is achieved by the integration, development and use of the most up-to-date optical technologies to image the dynamic cellular composition of the pathological central nervous system in control and treatment conditions. We also aim at tuning optical methodologies for their use in combination with implantable stimulating/recording electronic devices for Human Machine Interface.

Equipments

  • Intravital Zeiss LSM 780 spectral two-photon excitation microscope coupled to a
  • Chameleon Ultrafast II femto second pulsed Infrared laser and
  • Coherent Optical Parametric Oscillator : multimodal multicolor 2P-CARS imaging
  • Surgery microscopes
  • Wetlab

Key ongoing projects

    • NEUROFIBRES : Design and characterization of a carbon based electrostimulable neuroprosthesis to promote axonal regeneration and functional recovery after spinal cord injury.
      • Coordinator : J. Collazos Castro ((National Hospital for Paraplegics of Toledo, Spain)
      • Participant : F. Debarbieux (INT, France), F. Kirchhoff (USAAR, Germany), A. Ferrari (UCAM, England), S. Achart (Axon’cable, France), N. Pugno (UNITN, Italy), A. Eriksson Karlström (KTH, Sweden)
    • NEUROINFLAMDYN : Characterization of the immune response in multicolor fluorescent mice model of multiple sclerosis using intravital two-photon microscopy and multiparametric flow cytometry.
      • Coordinator : F. Debarbieux (INT, France)
      • Participant : M. Malissen (CIML, France) / H. Luche (CIPHE, France)
    • MYDEEPCARS : Implementation of intravital in depth polarized CARS imaging to dynamically monitor pathological changes of myelin ultrastructure
      • Coordinator : S. Brasselet (Institut Fresnel, France)
      • Participant : F. Debarbieux (INT, France)
    • Effect of clinically used antiangiogenic drugs on the cellular composition of tumor microenvironment in a murine model of glioblastoma. A study combining intravital two-photon microscopy and multiparametric flow cytometry.
      • Coordinator: G. Rougon (INT, France)
      • Participant : O. Chinot (APHM, France) / D. Figarella (APHM, France)/ M. Malissen (CIML, France) / H. Luche (CIPHE, France) / F.Debarbieux (INT, France)
    • Identification of the main tumoral and stromal cellular targets responsible for the beneficial action of pro-apoptotic drugs in a murine model of glioblastoma.
      • Coordinator: A. Tchoghandjian (CRO2, France)
      • Participant : D. Figarella (APHM, France) / F.Debarbieux (INT, France) – G. Rougon (INT, France)
    • Studying the influence of microenvironment on the microtubule dynamics in glioblastoma tumor cells
      • Coordinator : S. Honoré (CRO2, France) – D. Braguer (CRO2, France)
      • Participant : F. Debarbieux (INT, France) – G. Rougon (INT, France)

Médias

Spinal cord lesion

Spinal lesion in a triple fluorescent mouse to reveal inflammation

3D rendering of the inflammed spinal cord of a EAE induced triple fluorescent mouse

Time lapse showing interaction between yellow microglia and green monocyte derived cells in the spinal cord of an EAE induced triple fluorescent mouse

References

    1. Gasecka P., Jaouen A., Bious F.Z., Barbosa de Aguiar H., Duboisset J., Ferrand P., Rigneault H., Balla N., Debarbieux F., Brasselet S. (2017) Degradation of molecular organization of myelin lipids in autoimmune demyelination probed by polarization resolved nonlinear vibrational microscopy. J. ISSN: 0006-3495 ; doi.org/10.1101/105965
    2. Ricard C., Lamasse L., Jaouen A., Debarbieux F. (2016) Combination of an optical parametric oscillator and quantum-dots 655 to improve imaging depth of vasculature by intravital multicolor two-photon microscopy Biomedical Optics express 7(6):2362-72. doi: 10.1364/BOE.7.002362.ISSN: 2156-7085
    3. Ricard C., Tchoghandjian A., Luche H., Grenot P., Figarella-Branger D., Rougon G., Malissen , Debarbieux F. (2016) Dynamic phenotype of microglial and monocyte-derived cells in glioblastoma bearing mice. Scientific Reports 19;6:26381. doi: 10.1038/srep26381. ISSN: 2045-2322
    4. Ricard C., Debarbieux F. (2014) Six-color intravital two-photon imaging of brain tumors and their dynamic microenvironment. Frontiers in Cellular Neuroscience. ISSN: 1662-5102.
    5. Ricard C., Stanchi F., Rodriguez T., Amoureux M.C., Rougon G., Debarbieux F. (2013) Dynamic quantitative intravital imaging of glioblastoma progression reveals a lack of correlation between tumor growth and blood vessel density PLoS One 8(9):e72655; eISSN-1932-6203
    6. Fenrich K.K., Weber P., Rougon G, Debarbieux F. (2013) Long and short term intravital imaging reveals differential spatiotemporal recruitment and function of myelomonocytic cells after spinal cord injury J Physiol. 591: 4895-4902.; ISSN : 0022-3751