MorphoMapping identifies redox-dependent control of NETotic states in primed neutrophils

Abstract Neutrophils rapidly deploy phagocytosis, degranulation, and neutrophil extracellular trap (NET) formation to control infections, yet exaggerated NET formation contributes to tissue injury in inflammatory disease. Because NETosis is tightly linked to the cellular redox environment, we developed MorphoMapping, an imaging flow cytometry–based pipeline that resolves neutrophil morphotypes at single-cell resolution using purely morphology-derived features. Applying MorphoMapping to cytokine-primed primary human neutrophils exposed to the heme-targeted redox modulator WF10 (tetrachlorodecaoxide) revealed a compact, treatment-specific morphological state and a reproducible redistribution of cells away from NETotic, H3Cit-associated morphotypes. Instead, WF10 enriched nuclear morphologies compatible with non-NETotic termination programs. These data indicate that defined heme-dependent redox modulation suppresses NETotic neutrophil states under priming conditions and establish MorphoMapping as a scalable approach to quantify redox-sensitive neutrophil state shifts in suspension.