Objective To evaluate the sclerosing effects of several chemical sclerosants on the gallbadder mucosa in vivo. Methods Twentyeight rabbits were randomly divided into： Normal saline was used as contrast agent, and 95% ethanol, 5% tetracycline and compound phenol as sclerosants respectively. The cystic duct was ligated and cholecystostomy was performed for each animal. Sclerosants and contrast agent were injected into the gallbladder through a tube respectively and were kept in the gallbladder for 15 minutes. Animals were killed two weeks later; the gallbladder, bile duct and adjacent liver tissue were obtained for histopathological examination. Results With the naked eye, normal saline and 5% tetracycline made the cavity of the gallbladder unchanged; 95% ethanol made two gallbladder cavities occluded, two smaller, and three unchanged; and compound phenol made four gallbladder cavities occluded, and three smaller. In histopathological observations, 95% ethanol and compound phenol could make seven rabbits’ gallbladder mucosa completely destroyed (7/7); compared with normal saline (1/7), there was significant difference (P＜0.01). Five percent tetracycline had some effect of ablating the gallbladder mucosa (5/7), but compared with normal saline, there was no difference (Pgt;0.05). Conclusion Ninety-five percent ethanol can be clinically applied as an effective and safe sclerosant.