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Mitochondria are vital organelles that not only produce cellular energy but also participate in many biological processes. Recently, various fluorescent probes have been developed for mitochondrial imaging. However, due to the lack of suitable dyes or strategies, it is difficult for most reported mitochondrial targeting probes to prove whether the analytes they detected are from mitochondria. In addition, positive charge on mitochondrial probes can seriously affect the mitochondrial environment. To address these issues, we herein put forward a novel strategy for probe design based on a smart NIR dye ( HDFL ) for mitochondrial targeting detection. Compared to general mitochondrial targeting probes that are modified with a target site and a reaction site, the new strategy is to combine the two sites together for a mitochondrial probe that would provide accurate detection of analytes in mitochondria without interference. As a proof of concept, we synthesized a mitochondrial-targetable probe HDFL-Cys for cysteine. Bioimaging studies have shown that the new type of probe HDFL-Cys can first accumulate in mitochondria and then react with the analyte (cysteine) accompanied by the departure of the targeting group (lipophilic cation moieties). Thus, it can specifically detect the analyte in mitochondria without interference from extra-mitochondrial analytes. We anticipate that the new strategy based on the novel NIR dye HDFL may be a potential platform for developing desirable ratiometric fluorescent probes for mitochondrial imaging.

Detection of analytes in mitochondria without interference from other sites based on an innovative ratiometric fluorophore