A fluorescent probe for lipid droplet polarity imaging with low viscosity crosstalk

Monitoring the variations of lipid droplet (LD) polarity is of great significance for the investigation of LD-related cellular metabolism and function. We hereby report a lipophilic fluorescent probe (BTHO) with the feature of intramolecular charge transfer (ICT) for imaging the LD polarity in living cells. BTHO exhibits an obvious attenuation of fluorescence emission in response to the increase of environmental polarity. The linear response range of BTHO to polarity (ε, the dielectric constant of solvents) is derived to be 2.21-24.40, and the fluorescence of BTHO in glyceryl trioleate falls in this range. Furthermore, BTHO has high molecular brightness, which may effectively improve the signal to noise ratio, along with the decrease of phototoxicity. BTHO exhibits excellent photostability and targeting capability to LDs with low cytotoxicity, which is satisfactory in long-term imaging in live cells. The probe was successfully applied for imaging LD polarity variation in live cells caused by oleic acid (OA), methyl-β-cyclodextrin (MβCD), H₂O₂, starvation, lipopolysaccharide (LPS), nystatin, and erastin. The low crosstalk caused by viscosity to BTHO measuring the LD polarity was confirmed from a calculation result.