The National Cytometry Platform (NCP) is an open facility accessible to all scientists at LIH as well as external researchers. The NCP is a shared resource facility that is available to assist researchers with cytometry experimental design, data acquisition, cell sorting and data analysis.


Activities

The National Cytometry Platform (NCP) within the LIH is part of the Department of Infection and Immunity and is headed by Dr Coralie Guerin.  It distributed over two LIH sites, the House of BioHealth in Esch sur Alzette and the BAM at the premises of the Centre Hospitalier Luxembourg.

It is state of the art technological platform benefiting from the latest technology, and of the expertise of qualified staff in flow cytometry, mass cytometry and imaging flow cytometry.

  • Flow Cytometry:

Flow cytometry is a powerful tool used to measure different aspects of a cell or particles in real time and high throughput. This technique allows to quantify mutiple cellular markers or processes, with the possibility of sorting subpopulation of interest. LIH’s NCP’s machines are able to measure up to 18 parameters at the same time.
Applications include immunophenotyping, cell viability, morphology, proliferation, secretion, cell cycle, microparticles quantification, rare events detection, endocytosis, phagocytosis, cell-cell interaction, in situ hybridization among other. These assays can utilize cell sorting technology, which physically isolates populations of interest for downstream applications including cell culture, molecular and cellular analysis, single cell cloning and many more.
Flow cytometry is adaptable and can be tailored to your particular inquiry; contact the core staff to discuss if this technology is the best choice for your research needs.

  • Mass cytometry:

Mass cytometry is a time-of-flight mass spectrometry based powerful technique use to perform deep immune profiling at a single cell level with up to 42 metal isotopes.

The NCP platform owns a Helios mass cytometer.  The range of isotopes (75 to 209 kD) allows the design of a 36 tagged-antibodies panel including extra- and intra- cellular markers, interleukines and immune checkpoints as well as DNA intercalant, live/dead discriminator and normalization beads.

The NCP’s team provides technical assistance (optimisation and antibody coupling) and expertise (panel design) for perfoming mass cytometric acquisition and analysis.

  • Imaging flow cytometry:

The ImageStream combines the speed and sensitivity of flow cytometry with the detailed imagery and structural information of microscopy thanks to high sensitivity CCD camera and is equipped with five lasers. An Image cytometer is able to capture high-resolution images of shape and fluorescence from cells in flow at rates of up to 1,000 cells per second.
Combination of flow cytometry high-throughput power with image resolution allows acquisition of statistically robust data on the detailed structure of cells and the location of intracellular components.

Equipment:

The cytometry core facility is equipped with two cell sorters, three analysers, one imaging flow cytometer, one mass cytometer and one microscope.
All machines are located in BSL-2 or BSL-3 areas.

  • Analyzers

BD ARIA SORP IIu
Lasers: 355 nm, 405 nm, 488 nm, 561 nm and 640 nm, 18 colors and 2 scatters

BD LSR Fortessa
Lasers: 355 nm, 405 nm, 488 nm, 561 nm and 640 nm, 18 colors and 2 scatters

BD Canto
Lasers: 488 nm and 633 nm, 6 colors and 2 scatters

  • Cell sorters

BD ARIA SORP IIu (BSL2)
Lasers: 355 nm, 405 nm, 488 nm, 561 nm and 640 nm, 18 colors and 2 scatters, 4 ways sorting
Collection into tubes (1.5 to 15 mL), plates (6 to 384 wells) or onto slides

BD Influx (BSL3)
Lasers: 355 nm, 405 nm, 488nm, 640 nm, 14 colors and 2 scatters, 6 ways sorting
Collection into tubes (1.5 to 15 mL), plates (6 to 384 wells)

  • Mass cytometer

Helios (https://www.fluidigm.com/products/helios#components)
Detection range 75 to 209 kDa

  • Flow imager

AMNIS Image stream MKII
Lasers: 405nm, 488nm, 561nm, 640 nm and 785 nm, 12 channels, 2 brightfields and 1 darkfield 20x, 40x, 60x objectives, extended depth of field (EDF)

  • Microscope

Zeiss Observer
Two cameras: black/white and color, 10x 20x and 63x objectives, apotome, LED with four filters set: 38HE, 43HE, 49 and 50

Links:

- 38 HE
- 43 HE
- 49
- 50

  • Softwares

Flow and mass cytometry: FlowJo, Kaluza, Diva, Modfit,Fcap, Cytobank
Imaging flow cytometry: Ideas, Inspire

Head of support service Unit

The NCP has been developed since 2004 and was continually updated the NCP with the state-of-the-art equipment. As of May 2015, a second cytometry site has been created in the House of BioHealth (HoBH). With acquiring the CyTOF2, BD LSRFortessa and the InFlux a strong structure has been created for the future scientific research in Luxembourg.

Featured Publications

Actin cytoskeleton remodeling drives breast cancer cell escape from natural killer-mediated cytotoxicity.

  • Cytoskeleton and Cancer Progression
  • National Cytometry Platform
  • NORLUX-Glioma Biology
  • Tumor Microenvironment
October 01, 2018
2018 Oct. Cancer Res.78(19):5631-5643. Epub 2018 Aug 13.
By:
  • Al Absi A
  • Wurzer H
  • Guerin C
  • Hoffmann C
  • Moreau F
  • Mao X
  • Brown-Clay J
  • Petrolli R
  • Casellas CP
  • Dieterle M
  • Thiery JP
  • Chouaib S
  • Berchem G
  • Janji B
  • Thomas C.

Intra-cardiac release of extracellular vesicles shapes inflammation following myocardial infarction.

  • National Cytometry Platform
June 22, 2018
2018 Jun. Circ Res.123(1):100-106. Epub 2018 Mar 28.
By:
  • Loyer X
  • Zlatanova I
  • Devue C
  • Yin M
  • Howangyin KY
  • Klaihmon P
  • Guerin CL
  • Kheloufi M
  • Vilar J
  • Zannis K
  • Fleischmann BK
  • Hwang DW
  • Park J
  • Lee H
  • Menasche P
  • Silvestre JS
  • Boulanger CM.
See all publications

Contact

Markus Ollert

M.D.

29, rue Henri Koch
L-4354 Esch-sur-Alzette
Luxembourg
Tel. : +352 26970-829

LOCATION

House of BioHealth
29 rue Henri Koch
L-4354 Esch-sur-Alzette
LUXEMBOURG

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Luxembourg - Bam
84, Val Fleuri
L-1526 Luxembourg,
LUXEMBOURG


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