Abstract: To meet the requirements of optical stability and biocompatibility for bioprobes, a comprehensive experiment on the synthesis, characterization, and bioimaging performance of nitrogen-doped carbon quantum dots (N-CDs) was designed. N-CDs were synthesized by a one-step hydrothermal method using yam powder and ethylenediamine as raw materials, both of which possess good biosafety. The morphology, structure and optical properties of the N-CDs were characterized and analyzed by transmission electron microscopy, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, UV-visible and fluorescence spectroscopy. Moreover, the biocompatibility and in vivo and in vitro imaging performance of the N-CDs were evaluated through cytotoxicity and bioimaging experiments. The experiment covers research content including material synthesis, structural characterization, biocompatibility evaluation, and bioimaging, which systematically demonstrates the complete process of scientific research. It helps cultivate students’ fundamental scientific thinking and improve their practical innovation ability.