Your name:
Your email:
Subject:
Your message:
CAPTCHA Image
To guard against spamming, thank you to fill the security code in the field below

The cytometry core facility of IFR94 pools, for research laboratories from Necker - Enfants Malades, two cytometers sorters and a flow cytometer Analyzer.
The cytometers are located on both sites, Necker and Broussais :

  NECKER BROUSSAIS
Referents =  Olivier Pellé Jérome Mégret Corinne Cordier
Cytometers =  BD FACSAria II SORP BD FACSAria I BD LSR Fortessa SORP

The Steering Committee is composed of the direction of the SFR and the members of the platform.

  • Jérôme Mégret  01 72 60 63 36 
    -  
  • Olivier Pellé  01 42 75 44 14 
    -  
  • Sorters are dedicated to sort class L1 and L2 cells, not containing any pathogens to humans or genetically modified viral vectors whose loss of pathogenicity is certainly established. Sorters are equipped with the automatic deposit unit: this option performs sorting and filing of a predefined number of cells on microscope slides, plates multi-well tissue culture and micro test-tubes for PCR strips.
    Sorts of bacteria and yeasts are not allowed on the Institute Imagine sorter.
    Class L2 cells must be fixed before any experimentation on the BD LSR Fortessa SORP.

    Présentation du BD FACSAria I (BD Biosciences)

    • 3 excitation lasers:
      • Laser Solid state 488nm
      • Laser Helium-Neon 632nm
      • Laser Solid state 407nm
    • Cell sorting with 1 to 10 fluorochromes
    • Sorting speed: 20 000 cells per second
    • Software: BD FACSDiva
    Configuration optique du BD FACSAria I
    BD FACSAria I optical configuration

    Présentation du BD FACSAria II SORP (BD Biosciences)

    • 5 excitation lasers:
      • Laser Solid state 488nm
      • Laser Helium-Neon 632nm
      • Laser Solid state 407nm
      • UV laser 355 nm
      • Laser 561 nm
    • Cell sorting with 1 to 16 fluorochromes
    • Sorting speed: 20 000 cells per second
    • Software: BD FACSDiva

    BD FACS Aria II SORP optical configuration

    Présentation du BD LSR Fortessa SORP (BD Biosciences)

    • 5 excitation lasers:
      • Laser Coherent Sapphire 488 nm
      • Laser Coherent Cube 639 nm
      • Laser Coherent Cube 405 nm
      • Laser Coherent Compass 561 nm
      • Laser Genesis 355nm
    • Detection: 17 parameters of fluorescence can be measured simultaneously
    • Software: BD FACSDiva


    BD LSR Fortessa SORP optical configuration

    Notes

    The Fortessa and the Imagine institut’s sorter have the same excitation lasers.
    These two machines have a laser for excitation to 561nm and a BD OctagonTM for the collection of signals (5 detected simultaneously).
    This laser allows the detection of red fluorescent proteins.
    It also allows to optimally excite the fluorochrome PE and all its derivatives tandem.
    Fluorochromes listed on the configurations above, are given indicatively, the lists are not exhaustive. Are absent markers of cell viability, cell functions and metabolism as well as other fluorochromes such as the Alexa Fluor.
    The platform staff is at your disposal to define with you the optimal combinations of fluorochromes for each device, depending on your project.

     

    Applications

    Characterization and isolation of cell populations under the following techniques:

    • phenotyping, multi-stain membrane and intracellular
    • analysis cells transfected with fluorescent reporter systems
    • analysis of hematopoietic stem cell "Side Population" (Hoechst 33342)
    • study of the mono-parametric cell cycle (permeabilized cells: IP, intact cells: Hoechst33342), study of the multi-parametric cell cycle (Incorporation of nucleoside: BrdU, EdU, marking of RNA with the Pyronin), study of the cell cycle with the colorful indicators (red and green) genetically encoded FUCCI
    • study of the vital parameters of the cell: cell proliferation (CFSE), mitochondrial membrane potential (JC-1, DiOC2, MitoTracker), enzymatic activity, membrane integrity (IP, dye Sytox, 7 - AAD), study of the signaling pathways (phospho-proteins)
    • multiplex assays (cytokines) with the BD Cytometric Bead Array
    • study of calcium flux (Indo-1)

    The service can develop new applications, on demand, on the basis of the technological potential of our equipment, and their possible evolution.

      NECKER BROUSSAIS
    Référents =  Olivier Pellé Jérome Mégret Corinne Cordier
    Appareils =  BD FACSAria II SORP BD FACSAria I BD LSR Fortessa SORP
    Adresse =  Bâtiment Pasteur – Porte P2
    Hôpital Necker – Enfants Malades
    149 rue de Sévres
    75015 Paris
    Bâtiment Leriche – Porte 9
    L-RB122

    Faculté de Médecine Paris Descartes
    96 rue Didot
    75014 Paris
    Bâtiment Leriche – Porte 9
    L-RB119

    Faculté de Médecine Paris Descartes
    96 rue Didot
    75014 Paris

     

    Proposed activities:

    • Realization of cell sorting in assisted sessions
    • Theoretical and practical training to become autonomous user (non-assisted sessions): training in the use of flow cytometers sorters and analysers and computer analysis tool (DivaSoftware)
    • Guidance, technical assistance, and development of experimental protocols
    • Assists in the interpretation of the results
    • Provision of the site FlowJo license


    PRICING

    Sessions assisted sorting: €25 / hour
    Non-assisted sessions sorting: €15 / hour
    Analyser: €14 / hour