Cell death plays a vital role in body development, maintenance of tissue function, and homeostasis. Accurate evaluation of cell death types is of great importance for pharmacological and pathological research. However, there is a lack of efficient fluorescent probes to discriminate various cell states. Here, we design and synthesize a novel activatable fluorescent probe PNE-Lyso to detect intracellular pH and hexosaminidases with two kinds of fluorescence signals. PNE-Lyso could distinguish dead cells from healthy cells based on a dual-color mode by targeting the lysosome and evaluating lysosomal hexosaminidase activity. Significantly, PNE-Lyso could also discriminate apoptotic and necrotic cells through visualizing lysosome morphology that is adjusted by the integrity of the lysosome membrane. Moreover, probe PNE-Lyso was successfully applied to investigate the drug-induced cell death process. To the best of our knowledge, this work is the first time cell death types have been distinguished based on a single fluorescent probe.

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