Objective: To investigate the effect of nitroglycerin on veinpuncture. Methods: 30 KM mice were divided in 3 groups at random. Group 1 was dealed with nitroglycerin, and the others were dealed with ethanol and distilled water respectively. Recorded the time and the degree which every mouse needed in every treatment, and the treatment period was from finding the puncture point to achieving the puncture. Results: Comparing with the others, nitroglycerin reduced the time and the degree of the vein puncture, the difference among groups was significant in late period. Conclusions: Nitroglycerin might ease venepuncture and improve operational efficiency.