Tetraphenylethylene-based AIE nanoprobes for labeling lysosome by two-photon imaging in living cells

Lysosomes are essential cellular organelles, serving vital functions in cellular metabolism and degradation. The design of specifically targeting lysosomes probes with aggregation-induced emission (AIE) characteristics using two-photon excitation techniques is significance and challenging work. Here we designed and synthesized two tetraphenylethylene (TPE)-based AIE fluorescence probes, naming TPE-Ma and TPE-Py, with TPE as the matrix and morpholine (Ma) or pyrrolidone (Py) as the targeting group. These probes exhibit a significant Stokes shift, low cytotoxicity, two-photo fluorescence imaging and lysosome-specific targeting capability ensuring their suitability for fluorescence imaging applications. To enhance the water solubility and cellular accumulation of TPE-Ma and TPE-Py in tumor cells, we employed a biocompatible polymer 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-mPEG₂₀₀₀) as a nanocarrier. By encapsulating TPE-Ma and TPE-Py within DSPE-mPEG₂₀₀₀, we successfully developed two AIE fluorescent nanoprobes known as DSPE@ TPE-Ma and DSPE@ TPE-Py. The results demonstrated that fluorescent nanoprobes DSPE@ TPE-Ma and DSPE@ TPE-Py possess excellent cell permeability, biocompatibility, superior photostability and specific targeting towards lysosomes in MCF-7 cells. Our findings highlight the potential of these fluorescent nanoprobes as effective tools for two-photon fluorescence imaging and targeted detection of lysosomes in cancer cells.