To explore a method to rapidly differentiate between the sensory and motor fascicles in peripheral nerves for clinical application. Methods: The Sunderland V injury model was induced in the bilateral femoral nerve trunk, the muscle branch of the femoral nerve and the saphenous nerve of Sprague-Dawley (SD) rats. The Annexin V-Carbon quantum dots (CDs) antibody complex was prepared and applied to the severed nerve ends. Fluorescence was assessed under a 380 nm UV light source at 5, 10, 15 and 20 minutes after staining under a fluorescence microscope. Results: The fluorescence intensity at 5 minutes was insufficient to fully display nerve bundles in the injured bilateral femoral nerve trunk, the muscle branch of the femoral nerve, and the saphenous nerve of SD rats. However, the intensity of staining increased significantly between 10 to 15 minutes, with the sensory fascicles of the femoral nerve trunk and saphenous nerve showing blue fluorescence upon Annexin V-CDs antibody complex staining, while no fluorescence was observed in the motor fascicles of the femoral nerve trunk or the muscle branch of the femoral nerve. The fluorescence intensity gradually decreased after 20 minutes. There were no significant differences in the staining intensity within each group at 5, 10, 15 or 20 minutes. Conclusion: The Annexin V-CDs antibody complex can be used to effectively distinguish sensory from motor fascicles in peripheral nerves intraoperatively in SD rats.