Drug-induced liver injury (DILI) is considered gradually as a serious public health issue, and hepatotoxicity has been regarded as the main clinical problem caused by it. We suspected that both the intracellular viscosity and peroxynitrite (ONOO–) levels in drug-induced hepatotoxicity tissue are higher than those in a healthy liver. For this reason, we have presented a fluorescent probe VO for multichannel imaging viscosity and ONOO– simultaneously. Experimental results showed that VO has satisfactory detection performance for both viscosity and ONOO–, and based on the advantages of its lower cytotoxicity and pH-stabilities, VO was successfully employed to image viscosity and ONOO– in living cells and animals. More importantly, we use the probe to successfully showcase drug-induced hepatotoxicity by imaging viscosity and ONOO– induced by acetaminophen (APAP). All the results indicate that VO has great potential for the detection of viscosity and ONOO– and to assay drug-induced hepatotoxicity. Overall, this work offers a new detection tool/method for a deeper understanding of drug-induced organism injury.

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