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Examining the distribution and activity of lysosomes is important to elucidate the mechanisms of human diseases. Herein, we developed a novel approach to improve the visualization of lysosomes using lysosome-targeted fluorescent silica nanoparticles (Lys-SiNPs) for fast and reliable imaging of lysosomal integrity. Lys-SiNPs were synthesized through a one-pot process and showed excellent colloidal stability and desirable biocompatibility. Lys-SiNPs exhibited distinct fluorescence emission, high brightness, large Stoke shift and excellent photostability, owing to the coexistence of negatively charged mesoporous silica and fluorescent dye. Furthermore, Lys-SiNPs showed a lower cytotoxicity and faster cellular uptake than Cy5.5-SiNPs in HeLa and A549 cells. Importantly, Lys-SiNPs exhibited fast lysosomal sequestration and good cell permeability under the low pH environment, which were determined by lysosome imaging and in vitro cellular uptake experiments, respectively. In the lysosome stability assay, Lys-SiNPs be359ba680
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