Presentation

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The cytometry platform provides the research teams at the Necker site with the latest generation of cytometers. For all its cytometers, it offers assisted services, training so that users become autonomous (non-assisted services) as well as its expertise and assistance. The cytometry platform is also open to academic or private teams from other sites.

Location 

Plateforme de cytométrie de la SFR Necker

Faculté de Médecine Université de Paris

156-160 rue de Vaugirard

75015 Paris

 

Tél : 01 40 61 54 73

cyto.sfrnecker@inserm.fr

 

The SFR Necker cytometry platform is at 1st flour of the Faculty building

 

Staff

 
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Corinne Cordier

Head, IE Inserm​

corinne.cordier@inserm.fr

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Emmanuelle Six, PhD

Scientific referent​

emmanuelle.six@inserm.fr

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Simon Fillatreau, PhD

Scientific referent​

simon.fillatreau@inserm.fr

Jérome Mégret
Olivier Pellé
 

Equipements

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Sony SH800

 

3 lasers : 488, 561 and 640nm

6 fluorescences can be detected simultaneously

 

70, 100 and 130um nozzles

 

Sorting speed is 12,000 cells per second

Services

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The plateform will assist the non-autonomous users for their experimental adjustments and their experiments

 

Platform staff make their expertise and knowledge available to all users.

It assists users in the development of the experimental design :

combination of fluorescences, choice of experimental controls, sorting mode, experimental strategy.

 

Platform staff train users on all cytometers to become autonomous.

 

Applications

  • phenotyping, membrane and intracellular multi-labeling

  • analysis of transfected cells with fluorescent reporter systems (GFP, mCherry, etc.)

  • side population hematopoietic stem cell analysis (Hoechst 33342)

  • study of the mono-parametric cell cycle (Permeabilized cells: IP, Intact cells: Hoechst33342), study of the multi-parametric cell cycle (Incorporation of nucleosides: BrdU, EdU, RNA labeling with Pyronine), study of the cell cycle with colored indicators (red and green) genetically encoded FUCCI

  • analysis of cell death by apoptosis

  • study of the vital parameters of the cell: cell proliferation (CFSE), mitochondrial membrane potential (JC-1, DiOC2, MitoTracker), enzymatic activity, membrane integrity (PI, Sytox dyes, 7-AAD), study of signaling pathways (phosphoproteins)

  • multiplex assays (cytokines) with the BD Cytometric Bead Array

  • study of calcium flux (Indo-1) 

 

The platform can develop new applications according to the technological potential of the equipment and its possible evolution.

Publications

Direct contribution of skeletal muscle mesenchymal progenitors to bone repair. Julien A, Kanagalingam A, Martínez-Sarrà E, Megret J, Luka M, Ménager M, Relaix F, Colnot C. Nat Commun. 2021 May 17;12(1):2860.

 

Maturation and persistence of the anti-SARS-CoV-2 memory B cell response. Sokal A, Chappert P, Barba-Spaeth G, Roeser A, Fourati S, Azzaoui I, Vandenberghe A, Fernandez I, Meola A, Bouvier-Alias M, Crickx E, Beldi-Ferchiou A, Hue S, Languille L, Michel M, Baloul S, Noizat-Pirenne F, Luka M, Mégret J, Ménager M, Pawlotsky JM, Fillatreau S, Rey FA, Weill JC, Reynaud CA, Mahévas M. Cell. 2021 Mar 4;184(5):1201-1213.e14.

 

Toll-like receptor-9 stimulated plasmacytoid dendritic cell precursors suppress autoimmune neuroinflammation in a murine model of multiple sclerosis. Letscher H, Agbogan VA, Korniotis S, Gastineau P, Tejerina E, Gras C, Mégret J, Moe A, Drobyski WR, Zavala F. Sci Rep. 2021 Feb 26;11(1):4735.

 

Hepatitis B virus X protein promotes DNA damage propagation through disruption of liver polyploidization and enhances hepatocellular carcinoma initiation. Ahodantin J, Bou-Nader M, Cordier C, Mégret J, Soussan P, Desdouets C, Kremsdorf D. Oncogene. 2019 Apr;38(14):2645-2657.

 

A splenic IgM memory subset with antibacterial specificities is sustained from persistent mucosal responses. Le Gallou S, Zhou Z, Thai LH, Fritzen R, de Los Aires AV, Mégret J, Yu P, Kitamura D, Bille E, Tros F, Nassif X, Charbit A, Weller S, Weill JC, Reynaud CA. J Exp Med. 2018 Aug 6;215(8):2035-2053.

 

BAFF and CD4+ T cells are major survival factors for long-lived splenic plasma cells in a B-cell-depletion context. Thai LH, Le Gallou S, Robbins A, Crickx E, Fadeev T, Zhou Z, Cagnard N, Mégret J, Bole C, Weill JC, Reynaud CA, Mahévas M. Blood. 2018 Apr 5;131(14):1545-1555.

 

Periosteum contains skeletal stem cells with high bone regenerative potential controlled by Periostin. Duchamp de Lageneste O, Julien A, Abou-Khalil R, Frangi G, Carvalho C, Cagnard N, Cordier C, Conway SJ, Colnot C. Nat Commun. 2018 Feb 22;9(1):773.

 

Treatment of ongoing autoimmune encephalomyelitis with activated B-cell progenitors maturing into regulatory B cells. Korniotis S, Gras C, Letscher H, Montandon R, Mégret J, Siegert S, Ezine S, Fallon PG, Luther SA, Fillatreau S, Zavala F. Nat Commun. 2016 Jul 11;7:12134.

 

A unique CD8(+) T lymphocyte signature in pediatric type 1 diabetes.

Hamel Y, Mauvais FX, Pham HP, Kratzer R, Marchi C, Barilleau É, Waeckel-Enée E, Arnoux JB, Hartemann A, Cordier C, Mégret J, Rocha B, de Lonlay P, Beltrand J, Six A, Robert JJ, van Endert P. J Autoimmun. 2016 Sep;73:54-63.

 

Lymphoid Gene Upregulation on Circulating Progenitors Participates in Their T-Lineage Commitment. Zepponi V, Michaels Lopez V, Martinez-Cingolani C, Boudil A, Pasqualetto V, Skhiri L, Gautreau L, Legrand A, Megret J, Zavala F, Ezine S. J Immunol. 2015 Jul 1;195(1):156-65.

 

Innate pro-B-cell progenitors protect against type 1 diabetes by regulating autoimmune effector T cells. Montandon R, Korniotis S, Layseca-Espinosa E, Gras C, Mégret J, Ezine S, Dy M, Zavala F. Proc Natl Acad Sci U S A. 2013 Jun 11;110(24):E2199-208.