All-in-One Fluorescent Probe: Enabling Simultaneous Tracking of Multiple Dynamic Organellar Perturbations during Ferroptosis

lights up mitochondria/lysosomes (∼719 nm) and LDs (∼425 nm) simultaneously with high fidelity at distinctly separated emission wavelengths. Notably, mitochondria and lysosomes can be effectively distinguished by their distinct morphological features and fluorescence lifetimes, while changes in viscosity within mitochondria/lysosomes can be further quantified based on lifetime variations. Using this multifunctional probe, we visualized the dynamic process of Erastin-induced ferroptosis: LD accumulation, increased lysosomal viscosity, and decreased mitochondrial viscosity. Furthermore, we demonstrated that SLC7A11, a key regulatory factor of ferroptosis, restores the normal morphology and viscosity homeostasis of these organelles, highlighting the critical role of maintaining subcellular organellar morphology and microenvironmental stability in resisting ferroptosis.