OMIP-0XX: 40-Color Spectral Flow Cytometry Delineates All Major Leukocyte Populations in Murine Lymphoid Tissues

We designed this panel to deeply immunophenotype major leukocyte populations in primary and secondary murine lymphoid tissues as a function of time and treatment. This panel uses a robust set of extracellular markers capable of differentiating leukocyte subsets without the use of intracellular staining, thus allowing for the use of cells in downstream functional experiments or multiomic analyses. Our panel classifies T cells, B cells, natural killer cells, innate lymphoid cells, monocytes, macrophages, dendritic cells, basophils, neutrophils, eosinophils, progenitors, and their functional subsets by using a series of co-stimulatory, checkpoint, activation, migration, and maturation markers. Importantly, this panel can be used in pre-clinical settings to understand how diseases and corresponding treatments modulate leukocyte abundance and/or function in a systems wide setting.
Conclusion:
Our 40-color panel performs well in a variety of lymphoid tissues and discriminates all major leukocyte populations with excellent resolution. As such, this panel fills the niche for a modern day murine immunophenotyping tool that can be used in pre-clinical settings. Prior to data acquisition, we ensured that our Cytek Aurora cytometer was performing optimally by running a daily QC with SpectroFlo QC beads. All checks passed.