Flow Cytometry Analysis 

Flow Cytometry provides rapid qualitative and quantitative analysis of multiple physical and chemical characteristics and properties of single cells. It allows multiparametric analysis that can provide cell counts and measure cell size, cell granularity, density and expression levels of surface and intra-celllular molecules. It can be used, for example, to differentiate cell types in a heterogeneous cell population, determine the extent of purity of isolated subpopulations, assess the activation levels of the cell subpopulations and determine the ratios between various cell types..

IMMUNI T currently holds 2 Beckman's Cytoflex cytometers providing high-throughput sample handling capacity and versatility in the applications. We process all sample types from human or animal tissue, whole blood, fresh or frozen PBMC or cell lines.

Technical parameters: the ultra-low pressure peristaltic sheath and sample delivery system allow volumetric measurements with continuous flow. The absolute count in each analyzed population is also possible without using counting beads. Moreover, the use of Avalanche Photodiode detectors provides exceptional sensitivity across the entire visible spectrum. Finally, this cytometer displays 7-decade dynamic range and for the multiplex analysis of 15 parameters.

 

Our expertise and ability in flow cytometry together with our strength in designing, developing and performing customized cell-based assays provide a unique and specialized service offering.

Immunophenotyping

IMMUNI T provides an array of panels for phenotyping various cell populations. All the available panels can be customized and new panels can be designed based upon your needs.
Pre-designed panels are available for the phenotyping of the following cell populations:

  • T lymphocytes: CD4 subsets (Treg, Th1, Th2, Th17), CD8 subsets

  • B lymphocytes: B cells, plasma cells, Breg

  • NK/NKT cells

  • Monocytes, granulocytes and dendritic cells

  • MDSC (Myeloid Derived Suppressor Cells)

Cell counts, enumeration, ratios of biomarkers of interest, percent expression to mother gate are determined for each panel analyzed. The extent of analysis and the form of presentation of the data are performed as desired.

Immunoprofiling, Biomarker analysis

IMMUNI T provides various solutions for assessing the functional state of immune cells, establishing the desired profiles of cell populations as desired. Activation status, immune checkpoints, phosphorylation state of intracellular molecules, cytokine secretion with both intra- and extra-cellular measurements, cell degranulation and proliferation state can be assessed. These measurements lead the way to biomarker discovery/assessment in response to a particular treatment.

The following capabilities are offered:

  • Intracellular staining

    • Phosphorylated/unphosphorylated status of transcription factors or kinases

    • Expression level of intracellular cytokines and proteins

  • Peptide/protein detection and measurements

    • Multiplex cytokine measurements on CBA platform (Cytometric Bead Array)

    • Cell surface marker density

      • Cell identity

      • Activation status

  • Proliferation and viability

    • CFSE, CTV, eFluor670, Ki-67 staining

    • 7-AAD and other viability dyes

Detection of T lymphocyte proliferation by CTV (Cell Trace Violet) in vitro
PBMCs from a healthy donor were labelled with CTV and cultured for 6 days in the presence of PHA (top), a polyclonal stimulus that activates both CD4+ and CD8+ T cells, or with no stimulus (bottom). Proliferation is observed as a dilution pattern of the CTV in stimulated cells whereas no cell division is observed in unstimulated cells.

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Telephone : +1(514) 545 3097 

Fax : +1(514) 545 3064

Email : bud@immunit.com