Flow Cytometry Services
Reaction Biology’s team of flow cytometry experts build on many years of experience in performing flow cytometry analysis as a readout for in vivo studies as well as in vitro applications. Our scientists have access to a suite of state-of-the-art flow cytometers and sorters with various laser setups and can multiplex up to 20 markers for both, cell analysis and cell sorting.
Flow cytometry enables the identification of individual cells in a cell population, such as a tumor cell suspension, by staining 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 studies can be performed as endpoint readout of in vivo efficacy studies to investigate immune-modulating drug candidates.
Expression analysis of proteins can be performed with tumor material, blood, cell culture, or other samples for biomarker analysis. Our scientists will help you to determine the best suitable marker panel for your research project.
We offer four staining panels for immediate immune profiling of tumor samples. Custom-tailored panels can be established upon request tailored to the specific needs of your target protein or immune cell population of interest.
- Four staining panels are immediately available for immune phenotyping of tumor material
- Customizable panels are an option
- A suite of state-of-the-art flow cytometers and cell sorters
- Up to 20 markers can be multiplexed
Instrumentation
| Flow cytometer Instrument | Laser | Marker | Comments |
|---|---|---|---|
| BD LSR Fortessa | 4 | 17 | |
| Guava easyCyte 8HT | 2 | 8 | |
| Sony SP6800ZE Spectral Analyzer | 3 | 20 | full spectrum analyzer uses spectral unmixing instead of conventional compensation |
| Cell sorter Instrument | Laser | Marker | Comments |
|---|---|---|---|
| BD FACS Aria Fusion | 4 | 18 | Sort 4 cell populations simultaneously |
| BD FACS Aria III | 5 | 21 | Sort 4 cell populations simultaneously |
| Beckman Coulter MoFlo Astrios EQ | 4 | 21 | Sort 6 cell populations simultaneously |
Established staining panels
| Markers: |
| CD4+ T cells |
| CD8+ T cells |
| Treg cells |
| NK cells |
| B cells |
| M-MDSCs |
| PMN-MDCSs |
| DCs |
| M1 / M2 Macrophages |
| Neutrophils/ Granulocytes |
| B cells |
| Markers: | Memory Status: | Activation Markers: | ||
| CD4+ T cells | CD62L | PD-1 | ||
| CD8+ T cells | CD44 | Ki67 | ||
| Treg cells | CD122 | |||
| NK cells | ||||
| NKT cells |
| Markers: | Activation Markers: | |
| CD4+ T cells | Granzyme B | |
| CD8+ T cells | IFN-gamma | |
| Treg cells | CD107a (lamp 1) | |
| CD69 |
| Markers: | Additional Markers: | |
| T cells | PD-L1 | |
| B cells | CD86 | |
| Macrophages | ||
| cDC1 | ||
| cDC2 | ||
| pDC |
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 established, ready-to-use staining panels that allow for the analysis of various immune cell populations. In addition, we can custom-tailor our existing panels or establish new antibody panels to match our customer's research needs.
Method: Primary tumor material is disrupted, erythrocytes removed, and up to 3x106 obtained single cells/well are dispensed. The single cells are stained with marker-specific fluorescent antibodies and analyzed by flow cytometry.
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: We can perform cytokine and chemokine quantification with our MESO Quickplex SQ120 assay platform. Please inquire for more information.
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