Flow Cytometry Applications
Flow cytometry is a versatile tool that efficiently allows the investigation of cells after compound treatment based on changes in protein expression, protein localization, or protein modifications. For example, flow cytometry is used to quantify the activity of drug candidates via quantification of substrate modifications, including phosphorylation. Other cellular processes such as apoptosis or cell signaling can also be measured.
At Reaction Biology, flow cytometry is with cell culture, primary cells, tissue material, or blood. Our scientists have access to a full suite of state-of-the-art flow cytometers and cell sorters. Our team can further analyze sorted cells such as qPCR or immunofluorescence.
Our services include:
- Single-plex or multi-plex capacity with up to 20 markers
- Quantification of drug treatment effects on a cellular level
- Custom-tailored project design and assay development
- Samples can be created in-house by our cell assay or animal testing teams
Information about flow cytometry of tumor material for immunophenotyping can be found here.
Reach out today to talk to our scientist to find out how we can help you achieve your research goals and find out which methods are best suited to monitor the effects of your drug candidate.
Instrumentation - Flow cytometers and Cell sorters
| Flow cytometer Instrument | Laser | Marker | Comments |
|---|---|---|---|
| BD LSR Fortessa | 4 | 17 | |
| Cytoflex S | 3 | 9 | |
| 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 |
Examples of Flow Cytometry Applications
Investigating the expression level of the target protein after tumor treatment
Human JIMT-1 breast cancer tumors were grown in mice treated with vehicle or test substance. At the study endpoint, tumors were isolated and tumor material was disrupted with gentleMACS C tubes containers and the Tumor Dissociation Kit (Miltenyi Biotec) before antibody staining and flow cytometry analysis.
Gating: Single cells were selected for live events via live/dead dye. Human tumor cells were identified as negative for mouse CD45 marker and investigated for target expression on the cell surface. Untreated tumor cells served as positive control expressing the target on 99% of cells. Whereas only 14% of the tumor cells from treated animals were stained positive for the target.
In conclusion, treatment of JIMT-1 breast cancer with the test agent reduced the number of target expression tumor cells to 14%.
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