Suppressing ROS Production of AIE Nanoprobes by Simple Matrices Optimization for CNS Cell Observation and Minimized Influence of Cytoskeleton Morphology.

The visualization of the central nervous system (CNS) has proposed stringent criteria for fluorescent probes, as the inevitable production of reactive oxygen species (ROS) or heat generated from most photoluminescent probes upon excitation can disturb the normal status of relatively delicate CNS cells. In this work, a red-emitting fluorogen with aggregation-induced emission (AIE) characteristics, known as DTF, was chosen as the model fluorogen to investigate whether the side effects of ROS and heat could be suppressed through easy-to-operate processes. Specifically, DTF was encapsulated with different amphiphilic matrices to yield AIE nanoprobes, and their photoluminescent properties, ROS production, and photothermal conversion rates were examined. BSA@DTF NPs possessed 1.3-fold brightness compared to that of DSPE-PEG@DTF NPs and F127@DTF NPs but its ROS generation efficiency is markedly decreased to only 2.4% of that produced by F127@DTF NPs. Meanwhile, BSA@DTF NPs showed a negligible photothermal effect. These features make BSA@DTF NPs favorable for long-term live cell imaging, particularly for fluorescent imaging of CNS cells. BSA@DTF NPs were able to sustain the normal state of HT-22 neuronal cells with continuous illumination for at least 25 min, and they also preserved the cytoskeleton of microglia BV-2 cells as the untreated control group. This work represents a successful but easy-to-operate process to suppress the ROS generation of red-emissive AIEgen, and it highlights the importance of minimizing the ROS generation of the fluorescent probes, particularly in the application of long-term imaging of CNS cells.