NIR-fluorescent ceramide is a neutral near-infrared ( NIF ) fluorescent probe ( λ em =650 nm) based on the C-bridged nitrobenzoxadiazole SCOTfluor core, which enables visualization of lipid metabolic processes such as sphingolipid trafficking. The emission signal of NIR-fluorescent ceramide increases significantly in liposomal environments. NIR-fluorescent ceramide acts as an intracellular trafficking tracer, localizing to regions surrounding the endoplasmic reticulum and Golgi apparatus within a short period, and then translocating to recycling lysosomes over a longer period, thus enabling real-time visualization of sphingolipid trafficking and biological lipid functions in living cells. NIR-fluorescent ceramide also generates metabolic uptake profiles and provides multiple metabolic readouts in human cancer cell lines.
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