Cellular Immunophenotyping

With only small amounts of blood, peripheral blood mononuclear cells (PBMCs) or dissociated tissue a broad spectrum of cell types can be quantitatively and qualitatively analyzed. With only one sample a variety of lymphocyte subsets can be measured, using up to 13 colors (fluorophores) simultaneously.

We have a range of different standardized panels for analyses. Our basic panels include all major immunological cell types and assess additional phenotypic information for these cell subsets. For T cells, B cells and dendritic cells we have panels to distinguish different subtypes. For example: T cells can be divided into T regulatory cells, Th1, Th2, Th17 and TFh cells, each with its own effector cytokines (Lymphocyte cytokine profiling) and differentiation phenotypes. We also have panels specially designed to characterize leukemic and lymphoma cells and different stem and progenitor cells (MSCs, HSPCs, etc.).

In addition, we can custom design panels. The analyses of the data can be performed by gating populations of interest or by means of unsupervised clustering.  Unsupervised clustering can provide new insights in your data and may elucidate new cell subsets of interest. Setting up and measuring a panel to analyze in an unsupervised way is offered in co-development.

The design of the antibody-panels is based on the guidelines from the Dutch Society for Cytometry (NVC). The NVC is part the scientific partner of the section Immunological and Molecular Cell diagnostics of the Dutch Foundation for Quality Assessment in Clinical Laboratories. This foundation organizes external quality assessments which the laboratory for Immunocytology of Sanquin partly coordinates and in which Sanquin participates.

Basic phenotypical lymphocyte analysis

An example of a basic phenotypical lymphocyte analysis using healthy donor PBMCs. An overview of the cell populations is given including frequencies. Below three examples of density plots are shown. Left graph: Gated on CD3, CD4+ (T helper cells) and CD8+ (cytotoxic) populations. Middle graph: T helper cells have been divided into naïve, effector, central memory (CM) and effector memory (EM) cells using CCR7 and CD45R0. Right graph: Gated on CD3 and CD4 positive cells, T regulatory cells have been divined by CD25 and FoxP3.

Illustratie 4 400 pixels

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Last edited on: 1 December 2015