Cell Phenotyping

We provide full flow cytometry services and FACS services

Cell Phenotyping platform uses FACS - Fluorescence-Activated Cell Sorting - to support pre-clinical and clinical PoC studies in immunology, cancer research, stem cell studies, and various other fields where understanding and isolating specific cell populations is crucial.

Thanks to this platform it is possible to investigate the heterogeneity of cellular populations and gain insight into the role of different cell types in biological processes and diseases.

Cell Phenotyping Platform

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CELL PHENOTYPING

Full flow cytometry services (FACS services) from experimental design to specimen processing, acquisition (3 lasers 405 nm, 488nm, 640nm), and data analysis.
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Becton Dickinson FACSARIA III CELL SORTER

A brief review of our platform Cell Phenotyping

Cells

1. Assays

Cell Phenotyping

  • Analysis of extracellular markers, mono- and multi-parametric analysis (i.e. receptors, adhesion molecules, costimulatory molecules and MHC);
  • Analysis of intracellular markers and cytokines, chemokines and growth factors.

PBMC analysis of subpopulations

  • Th1, Th2, Th17, naive, Treg, memory, effector cells;
  • Simultaneous staining for intracellular cytokines and cellular populations.

Cell supernatant analysis

  • Multi-analyte detection: – Th1 Th2 Th17, inflammatory cytokines panels.

Cell sorting

Functional assays

  • cell signaling;
  • cell cycle;
  • degranulation
  • phosphorylation

2. Readouts

Cell Phenotyping

    CD62E expression on endothelial cells unstimulated (left panel) and stimulated with TNFa (middle panel). Histograms display PE-G586 fluorescence intensity before (in green) and after (in red) TNFa stimulation.

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    Intracellular Staining

    Interferon-g production in CD4+ lymphocytes upon stimulation with PMA/Ionomycin. Representative FACS plots of gating strategy for CD14- CD4+ cells (upper panels). Intracellular staining for Interferon-g in unstimulated cells (i), and PMA/Iono activated cells in absence (ii) or presence of immunomodulator (iii) (lower panels).

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    Extracellular vesicles

    1. Assays

    Microvesicle or Exosome Analysis

    • Qualitative:
      • Analysis of extracellular markers, mono- and multi-parametric analysis (i.e. receptors, adhesion molecules, costimulatory molecules and MHC);
      • Analysis of intracellular markers and cytokines, chemokines and growth factors.
    • Quantitative:
      • Quantitative analysis for characterization specific microvesicles-subsets.

    Microvesicle sorting

    2. Readouts

    Plasma EVs

      Plasma-derived and Tears-derived EVs stained with Violet 450 dye. In the left panels, the dimensional gate identifies EVs with 180-900 nm size. In the right panels, the gate shows fluorescently stained EVs population.

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      Tears EVs

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      Representative sorting layout for Plasma EVs

      Gating strategy for EVs sorting. Different EVs populations were identified by immunostaining for specific markers.

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