Leukocyte Subset Detection
18-Color PBMC Immunophenotyping: Ten Cell Types in One Run
Introduction
An 18-color panel is not simply a longer version of a four-color one. Spectral overlap, antibody titration and the gating strategy have to be settled together, and a panel that works on one instrument does not automatically work on the next.
Starting from a published panel removes most of that groundwork. This one builds on OMIP-024 (Moncunill et al., Cytometry Part A 2014, 85(12), 995–998), an optimized panel designed to read as much immune status as possible out of a small blood volume — originally for PBMC from children in vaccine research. It covers ten cell types, and it identifies regulatory T cells by CD25 and CD127, so no intracellular Foxp3 stain is needed.
What remains is the part that belongs to your own bench: matching the panel to the detector configuration of your instrument and re-titrating the antibodies on your own material. The application note shows that step carried out on the Agilent NovoCyte Quanteon, the instrument we supply and support — ask us for a demonstration if you would like to see the panel on your samples. It is marked for research use only and not for use in diagnostic procedures.
Understanding immune status requires precise monitoring of multiple leukocyte subsets. Using the Agilent NovoCyte Quanteon Flow Cytometer, researchers developed an 18-color immunophenotyping panel to simultaneously identify and quantify a wide range of cell types in human PBMCs — including monocytes, B cells, plasmablasts, CD4+ and CD8+ T cells, regulatory T cells, γδ T cells, NK T cells, NK cells, and dendritic cells.
Key Findings
This optimized panel, based on OMIP-024, enables:
Comprehensive immune profiling from limited sample volumes.
Accurate antibody titration with NovoExpress software to ensure optimal resolution and minimal background.
In-depth analysis of NK and T cell activation and differentiation, critical for evaluating immune responses to vaccines and infections.
Conclusion
The NovoCyte Quanteon empowers high-parameter, high-performance flow cytometry for broad and detailed immune surveillance. This 18-color panel demonstrates how modern cytometry can streamline complex analyses and expand our understanding of immune function.
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