Elucidating the intrinsic relationship between mitochondrial pH (pH m ) fluctuation and lipid droplets (LDs) formation is vital in cell physiology. The development of small-molecular fluorescent probes for discrimination and simultaneous visualization of pH m fluctuation toward LDs has not yet been reported. In this work, utilizing pH-driven polarityreversible hemicyanine and rhodamine derivatives, a multifunctional fluorescent probe is developed for selectively identifying mitochondria and LDs under specific pH values via dual-emission channels. This rapidresponse probe, Hcy-Rh, has two distinct chemical structures under acidic and alkaline circumstances. In acidic conditions, Hcy-Rh exhibits good hydrophilicity that can target mitochondria and display an intense red fluorescence. Conversely, the probe becomes lipophilic under weakly alkaline conditions and targets LDs, showing a strong blue emission. In this manner, Hcy-Rh can selectively label mitochondria and LDs, exhibiting red and blue fluorescence, respectively. Moreover, this ratiometric probe is applied to map pH m changes in living cells under the stimulus with FCCP, NAC, and H 2 O 2 . The interplay of LD− mitochondria under oleic acid treatment and starvation-induced autophagy has been studied using this probe at different pH values. In a word, Hcy-Rh is a potential candidate for further exploring mitochondria−LD interaction mechanisms under pH m fluctuation. Moreover, the polarity-dependent strategy is valuable for designing other functional biological probes in imaging multiple organelles.
The development of artificial blood substitutes based on hemoglobin (Hb) with high efficiency and biological safety is important for low blood oxygen tension or hypoxemia. A significant challenge is to...
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