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This study introduces a paradigm-shifting approach to optimize mitochondrial targeting. Employing a new fluorescent probe strategy, we unravel a combined influence of both Nernst potential (ψ) and partitioning (P) contributions. Through the synthesis of new benz[e]indolinium-derived probes, our findings redefine the landscape of mitochondrial localization by optimizing the efficacy of mitochondrial probe retention in primary cortical neurons undergoing normoxia and oxygen--glucose deprivation. This methodology not only advances our understanding of subcellular dynamics, but also holds promise for transformative applications in biomedical research and therapeutic development. [ABSTRACT FROM AUTHOR]

The dual-emissive fluorescent probe described here enabled dynamic tracking of singlet oxygen (1O2)-ROS species production and localization dynamics between the mitochondria and nucleoli in the presence of agents perturbing the mitochondrial membrane potential and under a photodynamic (PDT) system. Local structural information during the probe-1O2 interaction was followed by spectrally resolved confocal microscopy. [ABSTRACT FROM AUTHOR]