Presentation
The cytometry platform provides the scientific community with state-of-the-art analyzers and cell sorting cytometers. It also offers the expertise of its staff for:
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The development of experimental protocols and new applications in cytometry
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The design of multicolor panels and cytometric data analysis
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Training users in conventional and spectral cytometry, the use of cytometers, and data analysis (supervised or unsupervised)
The cytometry platform is open to all research institutes and private companies.
Flow cytometry is a technique that allows for rapid multiparametric analysis of individual suspended cells in a sample, which may consist of diverse or even extremely rare cell populations. The cell sorters can isolate specific cell subpopulations or deposit a single cell into each well of 96- or 384-well plates for cloning or single-cell analysis applications.
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
The SFR Necker cytometry platform is at 1st floor of the Faculty building
Staff
IE, Inserm
Johanna Soukaseum
AI-CDD
Equipements
The platform is equipped with 3 cell sorters and 2 analyzers.
It also offers a dedicated laboratory for biosafety level 2 infectious sample sorting by flow cytometry.
The Sony ID7000 spectral analyzer enables the simultaneous analysis of up to 41 fluorochromes per sample. It features an automatic sampler compatible with 96- and 384-well plates, 5 ml tubes, and a sample temperature control system.
The cell sorters are equipped with an automatic deposition system enabling the sorting and deposition of a defined number of cells into multi-well plates (96 or 384 wells), PCR micro-tube strips, or microscope slides. Single-cell sorting allows the deposition of one cell per well. The Index Sorting option precisely identifies each deposited cell.
Sony MA900 , dedicated to infectious sorting
4 lasers : 405, 488, 561 and 638nm
12 fluorescences can be detected simultaneously
70, 100 and 130um nozzles
Sorting speed is 12,000 cells per second
Sorter is confined under PSMII
Services
Assisted use: Support for experimental setup, multicolor panel design, and data analysis.
Unassisted use: User training on cytometer operation and data analysis for full autonomy.
The platform also provides access to several analysis software programs: Diva, Sony, FlowJo, Kaluza, and OMIQ, for multicolor data study using unsupervised analysis.
Cytometry Applications
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Phenotyping, multicolor surface and intracellular staining
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Analysis of transfected cells using fluorescent reporter systems (GFP, mCherry, etc.)
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Analysis of hematopoietic stem cells "Side Population" (Hoechst 33342)
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Cell cycle analysis (single-parameter with permeabilized or intact cells, multiparameter with nucleoside incorporation: BrdU, EdU, or RNA staining with Pyronin; or FUCCI-based indicators)
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Cell death/apoptosis analysis
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Analysis of cell vital parameters: proliferation (CFSE), mitochondrial membrane potential (JC-1, DiOC2, MitoTracker), enzymatic activity, membrane integrity (IP, Sytox dyes, 7-AAD), signaling pathways (phospho-proteins)
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Multiplex cytokine quantification (BD Cytometric Bead Array)
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Calcium flux analysis (Indo-1)
The platform can develop new applications depending on the technological capabilities and evolution potential of its equipment.
Publications
Severe hematopoietic stem cell inflammation compromises chronic granulomatous disease gene therapy.
Sobrino S., Magnani A.,Semeraro M., Martignetti L.,Cortal A., Denis A.,Couzin C., Picard C., Bustamante J., Magrin E., Joseph L., Roudaut C., Gabrion A., Soheili T., Cordier C.,
Lortholary O.,Lefrere F., Rieux–Laucat F., Casanova J. L.,Bodard S., Boddaert N., Thrasher A. J.,Touzot F., Taque S., Suarez F., Marcais A.,Guilloux A., Lagresle–Peyrou C., Galy A., Rausell A.,Blanche S., Cavazzana M., Six E.
Cell Rep Med 2023 Feb 21 ;4(2):100919
doi : 10.1016/j.xcrm.2023.100919
Mitochondrial dynamics and metabolic regulation control T cell fate in the thymus.
R. Elhage, M. Kelly, N. Goudin, J. Megret, A. Legrand, I. Nemazanyy, C. Patitucci,
V. Quellec, T. Wai, A. Hamaï, S. Ezine.
Front Immunol 2024 Jan 15:14:1270268
doi: 10.3389/fimmu.2023.1270268. eCollection 2023
Promitotic Action of Oenothera biennis on Senescent Human Dermal Fibroblasts.
S. Ceccacci, K. Roger, I. Metatl, C. Chhuon, K. Tighanimine, S. Fumagalli, A. De Lucia,
I. Pranke, C. Cordier, M. C. Monti and I. C. Guerrera.
Int. J. Mol. Sci. 2022, 23(23), 15153
doi : 10.3390/ijms232315153
Human anti-smallpox long-lived memory B cells are defined by dynamic interactions
in the splenic niche and long-lasting germinal center imprinting.
Chappert P, Huetz F, Espinasse MA, Chatonnet F, Pannetier L, Da Silva L, Goetz C, Mégret J, Sokal A, Crickx E, Nemazanyy I, Jung V, Guerrera C, Storck S, Mahévas M, Cosma A, Revy P, Fest T, Reynaud CA, Weill JC.
Immunity. 2022 Oct 11;55(10):1872-1890.e9
doi: 10.1016/j.immuni.2022.08.019
Analysis of mRNA vaccination-elicited RBD-specific memory B cells reveals strong
but incomplete immune escape of the SARS-CoV-2 Omicron variant.
Sokal A, Broketa M, Barba-Spaeth G, Meola A, Fernández I, Fourati S, Azzaoui I,
de La Selle A, Vandenberghe A, Roeser A, Bouvier-Alias M, Crickx E, Languille L, Michel M, Godeau B, Gallien S, Melica G, Nguyen Y, Zarrouk V,Canoui-Poitrine F, Noizat-Pirenne F, Mégret J, Pawlotsky JM, Fillatreau S, Simon-Lorière E, Weill JC, Reynaud CA, Rey FA, Bruhns P, Chappert P, Mahévas M.
Immunity. 2022 Jun 14; 55(6):1096-1104.e4
doi: 10.1016/j.immuni.2022.04.002
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.
doi: 10.1038/s41467-021-22842-5
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.
doi.org/10.1016/j.cell.2021.01.050
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.
doi: 10.1038/s41598-021-84023-0
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.
doi : 10.1038/s41388-018-0607-3
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.
doi: 10.1084/jem.20180977
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.
doi: 10.1182/blood-2017-06-789578
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.
doi: 10.1038/s41467-018-03124-z