Evaluation of intracellular lipid droplets viscosity by a probe with high fluorescence quantum yield

Background: Lipid droplets (LDs) are an important organelle as the main energy storage site in cells. LDs viscosity controls the material and energy exchange between it and other organelles. Furthermore, the LDs metabolic abnormalities, cell dysfunction, some diseases may be attributed to the singular LDs viscosity. Currently, the fluorescent probes for sensing the variations of LDs viscosity are still scarce and expose some drawbacks of low fluorescence quantum yield, low sensitivity and LDs polarity interference. Thus, the development of high performance probes is significant to detect LDs viscosity. Results: We hereby provide a lipophilic fluorescent probe (TPE-BET) with high fluorescence quantum yield (phi f, 0.91 in glycerol) for imaging LDs viscosity in living cells. With the increase of viscosity from 0.54 cp to 934 cp, the fluorescence at lambda ex/lambda em = 405/520 nm and the fluorescence quantum yield of TPE-BET linearly increased by 64.9 and 128.5 folds, respectively. Meanwhile, the outstanding LDs staining capability of TPE-BET may provide a high spatial resolution for LDs imaging. The cell imaging of TPE-BET not only successfully observed the viscosity variations of LDs in cell stress models, e.g., ferroptosis, inflammation and mitophagy, but also revealed the increased viscosity and extracellular delivery of LDs in heavy metal cell injury models (Hg/As) for the first time, which may supply concrete evidence for understanding the structure and function of LDs. Significance: This represents a new fluorescent probe TPE-BET with high fluorescence quantum yield for imaging LDs viscosity, which may decrease the dose of probe and excitation light intensity along with the improvement on signal noise ratio (S/N). The imaging results of TPE-BET clarified that LDs viscosity may be an appraisal index on cell differentiation, state evaluation and drug screening.