Ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1), a crucial biomarker on the surface of tumor cells, plays a significant role in immune suppression and cancer metastasis. In this study, we successfully developed a novel amphiphilic fluorescent probe, QM-AMP, based on quinoline-malononitrile aggregation-induced emission (AIE) characteristics, for the real-time, and specific detection of ENPP1 enzymatic activity in multiple cell types. QM-AMP exhibits excellent dispersibility in both hydrophilic and hydrophobic environments, effectively minimizing background fluorescence interference and ensuring high sensitivity in ENPP1 detection. Upon interaction with ENPP1, the AMP group of the probe is specifically recognized and cleaved by ENPP1, which triggers the cleavage of the O–P bond and results in an “off-on” AIE fluorescence signal. QM-AMP demonstrates favorable photostability and biocompatibility, showing clear differential responses across cells with varying ENPP1 expression levels. Additionally, it can monitor ENPP1 expression changes under different macrophage polarization states, providing a potential tool for investigating ENPP1's role in the tumor immune microenvironment. This study not only offers a new approach for in-situ detection of ENPP1 as a biomarker but also opens up new possibilities for early diagnosis and treatment of related diseases.

Image guided photodynamic therapy (PDT) combines fluorescence tracing and phototherapy, which can achieve a more accurate and effective treatment effect. However, traditional photosensitizers are limited by the aggregation-caused fluorescence quenching (ACQ) effect and low reactive oxygen species (ROS) generation in a hypoxic environment, resulting in poor imaging and treatment effect. Herein, we report a tricyano-methylene-pyridine (TCM)-based Type I aggregation-induced emission (AIE) photosensitizer (TCM-MBP), the strong electron acceptance (D-A) effect extends the wavelength to near-infrared (NIR) region to reduce the autofluorescence interference, and oxygen atoms provide lone pair electrons to enhance the inter system crossing (ISC) rate, thereby promoting the generation of more triplet states to produce ROS. The AIE photosensitizer TCM-MBP exhibited low oxygen dependence, NIR emission, and higher ROS production compared to commercially available Ce 6 and RB. After encapsulation with DSPE-PEG₂₀₀₀, TCM-MBP nanoparticles (TCM-MBP NPs) could penetrate to visualize cells and efficiently kill cancer cells upon light irradiation. This study provides an oxygen-independent AIE photosensitizer, which has great potential to replace the commercial ACQ photosensitizers.