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Graphene quantum dots (GQDs) are promising for bioimaging applications due to their unique optical properties and good biocompatibility. However, the facile synthesis of GQDs with long-wavelength emission and high quantum yield (QY) under long-wavelength excitation is still a challenge. Herein, dual-emissive GQDs were synthesized via a facile one-pot solvothermal method using 2,7-dihydroxynaphthalene (2,7-DHN) and piperazine anhydrous (PA) as the raw materials. The obtained GQDs exhibited a green-emission band (core state emission) and a red-emission band (surface state emission), which originated from the inner π-conjugated structure and various surface functional groups, respectively. The π-conjugated structure mainly comes from the aromatic rings in 2,7-DHN, and the N–H of PA acts as the main nitrogen source, which is beneficial for the QY. The obtained GQDs showed obvious concentration-dependent optical properties. Importantly, they are able to generate red emission with a high absolute QY of 54.9% in ethanol and 24.9% in water, among the best values of reported GQDs. In addition, the prepared GQDs have good water-solubility, low cytotoxicity, and superior stability against photobleaching. Based on the above advantages, the as-prepared GQDs were successfully applied for labeling HeLa cells in vitro. Therefore, our prepared GQDs demonstrated that they can be an efficient biomarker for fluorescent bioimaging. [ABSTRACT FROM AUTHOR]