Specific labeling of biomarkers with bright and high photostable fluorophores is vital in fluorescent imaging applications. Here, we report a general strategy to develop single-molecule dendritic nanodots with finely tunable optical properties for fluorescent imaging. The well-defined nanodots are based on the divergent growth of biodegradable polylysine dendrimers with a fluorophore as the core. By tuning the size and surface chemistry, we obtained fluorescent nanodots with excellent brightness and photostability, favorable pharmacokinetics, and multivalent tumor-targeting capability. The nanodots provided robust, stable, long-lasting, and specific fluorescence enhancement in tumor tissue with an tumor-to-normal ratio (TNR) of ∼3 and lasting over 5 days and an TNR up to ∼17, holding considerable promise for cancer imaging and image-guided surgery. This strategy significantly improves the performance of fluorophores and can be applied to other modality imaging probes.
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