Synchronous regulation of the photoluminescence and physicochemical characteristics of multicolor carbon dots (CDs) can fully realize their application potential in multicomponent imaging. Herein, by utilizing an acid-regulated synthetic strategy, green-emissive and orange-emissive CDs that target lipid droplets (LDs) and mitochondria (Mito) have been developed for fluorescence visualization of LD-Mito interactions. The finding of different molecular fluorophores reveals that the precursor undergoes different reaction pathways in neutral and acidic conditions, which alters the size of sp-conjugated domain and surface properties for the successful regulation of photoluminescence properties and organelle-targeting ability. Moreover, the one-step fabrication of these two CDs was also realized by lowering the dosage of acid. Therefore, the multicolor imaging of LDs and Mito has been achieved with one-step staining, disclosing that their interaction frequency decreases during the lipotoxicity process. This work successfully demonstrates the high coupling potential between multicolor CDs and organelle-interaction visualization, which would provide guidance on the correlation between photoluminescence features and other properties of multicolor CDs for extending application space.
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