24-color (30-marker) antibody panel for deep immunophenotyping of immune cells in human peripheral blood unmixed data

This OMIP provides a detailed description of a 24-color (30-marker) full spectrum flow cytometry panel to identify all major immune compartments of blood, neutrophils, basophils, eosinophils as well as T, B, MAIT, Treg, NK, NKT-like, gd T, monocytes and dendritic cells, basophils, ILCs and their associated subsets. Activation, differentiation, check point and senescence markers were employed to provide an in-depth immune profiling in a single sample.
Conclusion:
Full Spectrum Flow Cytometry allows detection of 30 markers simultaneously with good population resolution, allowing comprehensive in-depth immune cell profiling within a single biological sample.
Notes:
Unmixed data files Daily QC was performed using SpectroFlo® QC beads prior to acquiring samples to ensure that the cytometer was performing optimally. Daily QC assessed the instrument’s optical alignment and the system performance drift by measuring %rCVs and gains needed to place the beads at the target locations established for each detector. During the typical QC protocol, Laser delays and area scaling factors were also optimized, and gain settings adjusted to account for day-to-day instrument variability.