Abstract: Based on the orientation of “problem solving” and the latest research results, a comprehensive innovative experiment of chemical biology is reported. According to the principle of fluorescence resonance energy transfer (FRET), a ratiometric pH sensor was prepared by covalently lingking carbon quantum dots (CDs) with fluorescein isothiocyanate (FITC), which was applied to monitor the intracellular pH in real time. The experiments show that the cross and integration of the front fields including the novel nanomaterials synthesis, fluorescence spectrometric analysis, cell labeling and imaging. This experiment reflects the intersection and fusion of cutting-edge fields such as the synthesis of new nanomaterials, fluorescence spectrum analysis, cell labeling, and cell imaging. The experimental content is substantial and the results are stable and repeatable. In addition, the experiment covers the content for students to explore and practice independently, which helps to improve the students’ practical innovation ability.