Raman imaging of heme metabolism in situ in macrophages and Kupffer cells

Herein, we provide the Raman imaging results for different stages of erythrophagocytosis of senescent red blood cells executed by isolated murine primary Kupffer cells and a murine macrophage cell line (RAW 264.7). Images were recorded with the use of 488 and 532 nm excitation lines. The use of Resonance Raman spectroscopy allowed the visualization of the heme metabolism and tracking of the systemic iron recycling process inside isolated murine Kupffer cells and RAW.264 cells. Because of the application of the different experimental assays, the erythrophagocytosis in two types of cells was presented and associated with the presence of different forms of oxidized and degradative derivatives of hemoglobin species. Moreover, we observed an increase of lipid level and later formation of lipid droplets during the erythrophagocytosis process inside RAW 264.7 macrophages and murine Kupffer cells.