The cytometry core facility of SFR Necker pools, for research laboratories from Necker - Imagine and Broussais, devices dedicated to cytometry (sorters, analysers and Flow Imager), and the expertise of his staff.

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

• 	Corinne Cordier	01 72 60 63 34
  	Platform manager	View the contact form

	Jérôme Mégret	01 72 60 63 36
	-	View the contact form

	Olivier Pellé	01 42 75 44 14
	-	View the contact form

• 	Emmanuelle Six	01 42 75 44 06
  	Scientific officer	View the contact form

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.

BD FACSAria IIIu (BD Biosciences)

4 excitation lasers: Laser Solid state 488nm Laser Helium-Neon 632nm Laser Solid state 407nm Laser Solid State 561nm Cell sorting with 1 to 16 fluorochromes Sorting speed: 20 000 cells per second Nozzle 70, 85, 100 et 130um Software: BD FACSDiva 8 Index-Sorting

BD FACSAria I optical configuration

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 8 Index-Sorting

BD FACS Aria II SORP optical configuration

BD LSR Fortessa SORP (BD Biosciences)

SONY SH800 (Sorter Sony)

• 2 sources d’excitation :
  • Laser 488 nm
  • Laser 561 nm
  • Détection : 6 paramètres de fluorescence peuvent être mesurés simultanément

• Software : SONY SH800

SONY SP6800 (Spectral Analyser - SONY)

• 2 sources d’excitation :
  • Laser 488 nm
  • Laser 638 nm
  • Laser 405 nm
• Détection : Collecte des photons entre 400 et 800nm
• Software : SP6800

AMNIS Image Stream XMKII

• 5 sources d’excitation :
  • Laser 488 nm
  • Laser 642 nm
  • Laser 375 nm
  • Laser 405 nm
  • Laser 560 nm
• Software : Ideas

Choose the optimal combination of fluorochromes for your panels of antibodies is a complex process. The staff of the platform is at your disposal to define with you the combinations of fluorochromes for every device, according to 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.

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 2018 - sites Necker - Imagine - Broussais

For BD FACSAria IIIu, BD FACSAria II SORP, BD LSR Fortessa SORP, Sony SH800 and Sony SP6800 :

Autonomous user : 24 € / hour
Non - autonomous user : 35 € / hour

For the Imager in flow (AMNIS ImageStream) : see with michael.dussiot@institutimagine.org

The use of the posts of analysis is free.