Flow Cytometry Facility (FCF)

Flow Cytometry Facility (FCF)

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Flow cytometry is a technology that allows the analysis of cells, particles, or microorganisms as they pass through one or more lasers while suspended in a buffered saline solution. Each particle is analysed for visible light scattering and one or multiple fluorescence parameters; therefore, it must be possible to label the particles with one or more fluorescent substances (eg, fluorescence-conjugated antibodies, propidium iodide, or green fluorescent protein (GFP), etc). This multiparametric study allows analysing of subpopulations in complex samples.

The recently created IISPV Flow Cytometry Facility is dedicated to helping both internal and external IISPV scientists with technical-scientific advice to develop reliable flow cytometry assays that meet current quality standards.

With the collaboration of:

  • 01

    Acquisition and Analysis: Study of cells or particles based on their fluorescent or light scattering characteristics. Among the acquisition services offered are:

    • – Immunophenotyping
    • – Cell viability assays
    • – Apoptosis and cell cycle analysis
    • – Mitochondrial activation
    • – Fluorescent protein expression
    • – Multiparametric analysis of molecules in liquid solutions (cytokines)
  • 02

    Cell Sorting:  FACS Aria III is an analyzer and separator cytometer that allows cells or particles to be separated based on parameters previously defined by the operator.

  • 03

    Advice and experimental design: The Facility offers advice to users on experimental design, upon request of the principal investigator and according to the availability of the Unit’s staff.

  • 04

    Self-service: The self-service regime is targeted towards advanced users, previously trained and authorized by UCF.

  • 05

    Training: The Flow Cytometry Facility has started a series of training courses aimed at higher education and university students, specifically within the URV.

  • 06

    Analysis of Results: The Facility makes available to users a license of the FlowJo flow cytometry analysis program, which allows offline analysis of the results. In addition, the FACS Aria III and Cytek Northern Lights equipment includes the BD FACS DIVA™ and SpectroFlo analysis software, respectively.

· Flow cytometers:

  • – FACS Ària III (BD)
  • – Northern Lights (Cytek)

 

· Cell sorters

  • – FACS Ària III (BD)

 

· Magnetic cell sorter autoMACS

· Vortex, pipettes and refrigerator

 

  • 01

    FACS Aria III (BD) Cell Sorter

    The FACS Aria III (BD) is a high-speed, high-resolution particle and cell analyzer and separator flow cytometer. It is configured with 3 lasers (violet 405nm, blue 488nm and red 633nm) with the capacity to analyze 16 parameters in total: 14 fluorescences plus size (FSC) and complexity (SSC) (Equipment configuration).

    It allows the use of different pressures depending on the size and type of cell to be sorted. Once separated, the cells can be collected in 1.5 mL, 5 mL and 15 mL tubes. Cell sorting can be carried out in 1.5 mL, 5 mL and 15 mL tubes.

    Figure 1. FACS Aria III (BD Biosciences)
  • 02

    Flow Cytometer with spectral technology Northern Lights (Cytek)

    Northern Lights (Cytek) is a flow cytometer with 3 lasers (violet 405nm, blue 488nm and red 638nm), 41 detectors and 38 fluorescence channels. This spectral technology equipment makes it possible to collect all the light emission emitted by each fluorochrome, thus obtaining the spectral signature, which is the sum of the different spectral fingerprints united. This way, it allows greater flexibility in the design of antibody panels and a greater capacity to resolve cell populations.

    Figure 2. Northern Lights (Cytek)

Erika Scholz, PhD. Head of Flow Cytometry and Cell Sorting Facility.
A/e: erika.scholz@iispv.cat
https://orcid.org/0000-0002-7589-0517

 

Pilar Gutiérrez Ventas
A/e: pilar.gutierrez@iispv.cat
https://orcid.org/0000-0002-4761-4478

Oficina técnica IISPV / Planta 7 del Hospital Universitario Joan XXIII

Institut d’Investigació Sanitària Pere Virgili (IISPV)

Hospital Universitari Joan XXIII C/ Doctor Mallafrè Guasch, 4

43005 Tarragona

A/e: citometria@iispv.cat

Telèfon: +34 977 297 996