A 30-Color Full-Spectrum Flow Cytometry Panel to Characterize the Immune Cell Landscape in Spleen and Tumors within a Syngeneic MC-38 Murine Colon Carcinoma Model

Abstract

Purpose and Appropriate Sample TypesThis panel was designed to characterize the immune cell landscape in the mouse tumor micro-environment as well as mouse lymphoid tissues (e.g., spleen). Previous Optimized Multicolor Immunofluorescence Panels (OMIP) with conventional cytometry were examples of high-quality fluorescent-based flow cytometry panels to characterize either the T-cell compartment or the myeloid compartment [1, 2]. The advent of spectral cytometry has enabled sufficient markers to be included in a panel to comprehensively characterize the immune cell landscape including both the T-cell and the myeloid cell compartments. In this body of work, we demonstrated that we could measure the frequency and characterize the functional status of CD4 T cells, CD8 T cells, regulatory T cells, NK cells, B cells, macrophages, granulocytes, monocytes, & dendritic cells. This panel is especially useful for understanding the immune landscape in "cold" preclinical tumor models with very low immune cell infiltration, and for investigating how therapeutic treatments may modulate the immune landscape.