Objective: To compare the difference in laboratory indexes and imaging results among patients with gallstones in different locations, so as to provide reference for clinical diagnosis and differential diagnosis. Methods: The data of 146 patients with gallstones admitted from February 2011 to September 2013 were retrospectively analyzed. According to the location of stones, the patients were divided into gallbladder group (35 cases), intrahepatic group (49 cases) and extrahepatic group (62 cases), and the results of relevant laboratory indexes and imaging examinations among the three groups were compared and analyzed. Results: Liver function-related parameters: the levels of bilirubin and transaminases in gallbladder group were significantly lower than those in intrahepatic group and extrahepatic group, and the levels of alkaline phosphatase and γ-glutamyltransferase in intrahepatic group were significantly lower than those in gallbladder group and extrahepatic group (all P<0.05). Three routine tests: the number of white blood cells percentage of neutrophils in intrahepatic group were significantly lower than those in gallbladder group and extrahepatic group, and the urobilinogen level in in gallbladder group was significantly higher than that in intrahepatic group and extrahepatic group (all P<0.05); urine bilirubin test showed negative or weakly positive result in gallbladder group, and strongly positive result in intrahepatic and extrahepatic group; the color of stool in the gallbladder group was yellow, which was pale or clay-colored in intrahepatic and extrahepatic group. Imaging examinations: the detection rates of B-ultrasound and CT were relatively high in gallbladder group (88.6%, 91.4%), the detection rate of CT in intrahepatic group was relatively high (87.8%), and the detection rate of MRCP in extrahepatic group was relatively high (90.3%). Conclusion: The laboratory indexes and imaging results have certain reference value for diagnosis and differential diagnosis of gallstones in different locations, so a better understanding of their respective characteristics and complementary advantages may help in making a correct judgment.