Dead Cell Discrimination with Red Emissive Carbon Quantum Dots from the Medicinal and Edible Herb Echinophora tenuifolia

Abstract Accurately determining the viability of cells is crucial for in vitro cell research. Fluorescence-based live/dead cell staining is a highly desirable method to assess cell viability and survival in in vitro studies. We describe a green synthesis method to create red-emissive CQDs from the medicinal and edible herb Echinophora tenuifolia using microwave irradiation. We observed that the biocompatibility and photostability of the CQDs are superior. The antioxidant capacity of the CQDs and the plant extract were also investigated using different chemical methods (DPPH, ABTS, CUPRAC, FRAP, PBD, and MCA). The antioxidant capacity of the CQDs was similar to that of the extract of E. tenuifolia . Cytotoxicity studies indicate that while the CQDs are not toxic to L929, they exhibit significant toxicity towards HepG2 cells. The CQDs exhibited a strong negative zeta potential (-44.0 mV), which contributed to their selective interaction with dead cells while being repelled by viable cells with intact membrane potentials. The optimal concentration for effective, non-toxic imaging was determined to be 25 µg/mL, as lower concentrations did not produce detectable fluorescence. Differential staining experiments confirmed that CQDs selectively stained dead cells, with red fluorescence observed under the Texas Red filter. Moreover, CQDs exhibited favorable fluorescence intensity and stability, which may offer advantages for long-term and reliable bioimaging applications. In vitro studies on HepG2 and L929 cell lines revealed that the red-emissive CQDs from E. tenuifolia can be potentially used in bioimaging.