Protein Arginine Methyltransferase 5 (PRMT5) is an important player in breast cancer cell activity, and innovative fluorescent ligands targeting this enzyme offer revolutionary, real-time insights into its role in cancer progression, unlocking new avenues for diagnosis and treatment. This study introduces fluorescence-labeled PRMT5 ligands, highlighting their applications in visualizing PRMT5, monitoring enzymatic activity as well as studying toxicity. Herein, we describe the design, synthesis, and cellular imaging of a series of fluorescent ligands that target PRMT5. These ligands are based on the introduction of strong and selective PRMT5 inhibitors into various fluorophores using varied linkers. Among them, compound 7 at 10 μM was shown to exhibit strong fluorescence signals against MCF-7 cells with IC