Dual role far red fluorescent molecular rotor for decoding the plasma membrane and mitochondrial viscosity.

The dysfunctions in the mitochondria are associated with various pathological conditions like neurodegeneration, metabolic disorder, and cancer, leading to dysregulated cell death. Here, we have designed and synthesized a julolidine-based molecular rotor (JMT) to target mitochondria with far-red emission accounting for mitochondrial dysfunction. JMT showed viscosity sensitivity with 160-fold enhancement in fluorescence intensity. The origin of the dark state in a lower viscous environment was investigated through density functional calculations. We have employed JMT to monitor mitochondrial dysfunction induced by nystatin using confocal and fluorescence lifetime imaging microscopy. Further, we investigated mitochondrial abnormalities under inflammatory conditions triggered by lipopolysaccharide in live HeLa cells. The cellular uptake mechanisms of JMT were studied using various endocytosis inhibitors. Moreover, we reported tracking small fluorescent molecule switching from mitochondria to the plasma membrane upon introducing mitochondrial depolarizer in cells. On treating the mitochondria potential uncoupler, JMT relocates to the cell membrane and can be utilized for understanding the interplay between mitochondria and cell membranes. Moreover, JMT was applied to stain the RBC plasma membrane isolated from human blood.