Recent improvements in the flow cytometry technology allow the determination of the general immune status through the development of multicolor immunofluorescence panels. The one-tube multicolor flow cytometry assay (OTMA) that is presented here identifies 20 different, clinically relevant immune cell subsets and three common activation markers. Thereby, a comprehensive immune status that covers all major immune cells is easily obtained.The assay is suitable for every common three lasers and 10 color flow cytometer and includes the application of 15 different antibodies. Furthermore, the assay requires only 100 μL of EDTA-treated whole-blood and less than 40 min for sample preparation. By being easily adaptable to individual requirements and by additionally determining absolute cell counts, the assay is well-suited for translational research in clinical trials.
Keywords: Absolute cell count; Antibody; Gating strategy; Human whole-blood; Immune status; Immunophenotyping; Multicolor flow cytometry; One-tube measurement.