Flow Cytometry Services
Our flow cytometry capabilities allow the in-depth mode of action analysis of immune-modulating drugs.
Flow cytometry enables the identification of individual cells in a cell population such as a tumor cell suspension by staining against cell markers with antibodies conjugated with a fluorophore. The markers allow the distinguishment of a large variety of immune cell populations and their change in frequency upon treatment. Flow cytometry can also detect expression levels of proteins on cells for expression analysis of target proteins in distinct cell populations or biomarker analysis.
- 17 marker panel for a comprehensive investigation of tumor-infiltrating lymphocytes and other immune cell populations
- Customizable panels available
- A suite of state-of-the-art flow cytometers and cell sorters
Shown is the immune cell profile of one individual CT26wt tumor which was isolated at day 10 after tumor cell implantation into the mouse.
Primary tumor material is disrupted, erythrocytes removed, and up to 3x106 single cells/well dispensed. The single cells are stained with the marker panel and analyzed by flow cytometry using an LSR Fortessa (Becton Dickinson). The gating strategy is depicted below, starting at the large arrow in the middle.
| Marker | Immune cell population |
| CD45+; CD11b-; CD3e-; B220+ | B cells |
| CD45+; CD11b-; CD3e-; CD49b+ | NK cells |
| CD45+; CD11b-; CD3e+; CD8a+ | Cytotoxic T cells |
| CD45+; CD11b-; CD3e+; CD4+ | T helper cells |
| CD45+; CD11b-; CD3e+; CD4+; CD25+; Foxp3+ | CD4+ regulatory T cells |
| CD45+; CD11b+; F4/80+; CD206+; MHCII- | M2 macrophages |
| CD45+; CD11b+; F4/80+; CD206-; MHCII+ | M1 macrophages |
| CD45+; CD11b+; F4/80-; Ly6G+; Ly6C+ | Neutrophils/Granulocytes |
| CD45+; CD11b+; F4/80-; Ly6G-; Ly6C+ | MDCS |
| CD45+; CD11b+; F4/80-; Ly6G-; Ly6C-; CD11c+ | Dendritic cells |
Clinical data indicate that only a part of patients responds to treatment with immune-modulating drugs. It has hence been recognized as essential to consider the immune response already during preclinical drug development. Flow cytometry is widely used for the analysis of immune cell populations in tumors.
Panel: A comprehensive analysis of numerous immune cell populations in one tumor is a challenge. We use one staining panel that allows for the analysis of all major immune cell populations and comprises live/dead dye and antibodies against CD3, CD4, CD8a, CD45, CD25, CD11b, Ly6C, Ly6G, F4/80, CD11c, MHC class II, CD206, CD335, CD49b, B220, and FoxP3. The panel can be used for the analysis of tumor tissue and other organs.
Method: Primary tumor material was disrupted, erythrocytes removed, and up to 3x106 obtained single cells/well dispensed. The single cells are stained with the all-in-one marker panel and analyzed by flow cytometry using an LSR Fortessa (BectonDickinson). The gating strategy is depicted below, starting at the large arrow in the middle.
Report: All data of the mouse study, including tumor growth and flow cytometry data, will be presented in a comprehensive report, custom-tailored to the client study, and written by a Ph.D. level technical writer. The analysis enables you to draw conclusions directly from the study outcome and use the data for the filing of official documents.
Add-on cytokine quantification: Based on tumor tissue or blood we can perform cytokine and chemokine quantification with our MESO Quickplex SQ120 assay platform. Please inquire for more information.
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