A coumarin-based TICT fluorescent probe for real-time fluorescence lifetime imaging of mitochondrial viscosity and systemic inflammation in vivo.

Systemic inflammation, linked with abnormal mitochondrial viscosity, is reported to be associated with cerebro-cardiovascular disease and Alzheimer's disease. Therefore, it is of great significance to detect the mitochondrial viscosity to indicate the inflammatory signal in vivo. Considering the strategies of fluorescent molecular rotors (FMRs) and fluorescence lifetime imaging microscopy (FLIM), we have rationally designed a novel mitochondrial viscosity-specific fluorescent probe Mito-VCI, based on coumarin fluorophores with benzo[e]indolium as the rotor group. In a high viscosity solution system, the fluorescence lifetime of the probe Mito-VCI was prolonged due to the planarization and rigidity enhancement of the molecular rotor. Satisfactorily, the probe was only sensitive to viscosity, instead of non-viscosity factors such as pH and polarity. Furthermore, the probe sensitively targeted mitochondria in HeLa cells with a Pearson's correlation of 0.93, and specifically detected dynamics variation of mitochondrial viscosity with FLIM imaging in HeLa cells induced by LPS. Notably, significant fluorescence lifetime changes of Mito-VCI between normal and inflammatory tissues also occurred (for example, the fluorescence lifetime in the spleen changed from 1.128 to 1.432 ns). It can be inferred from the above observations that Mito-VCI could work as an effective and sensitive fluorescent molecular rotor for mitochondrial viscosity monitoring through FLIM imaging with a systemic inflammatory response, and provide potential applications for the diagnosis of systemic inflammation in pharmacology and toxicology studies.