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Background: Spillage of stones in peritoneal cavity is quite common in patients undergoing laparoscopic cholecystectomy.Lost stones may cause wide range of complications postoperatively which reflects the infective potential of these stones. Objective: The aim of this study was to find out the frequency of viable bacteria in stones retrieved from the gallbladder after laparoscopic cholecystectomy & subjecting the stones for culture. Design: Cross-sectional study. Place &Duration: This study was conducted at Surgical Unit 4 of Civil Hospital Karachi,from July 2013 till March 2014. Materials & Metods: 80 cases of cholelithiasis , selected & operated by laparoscopic cholecystectomy were included. During cholecystectomy , stones were collected , intact stones were sent in two separate containers from each subject, for microbiological examination. Results: This study included 80 patients, 58 patients(72.5%) were female & 22 patients (27.5%) were male. Out of these 80 patients, 27 patients (33.75%) had positive stone culture, out of these 27 patients , 16 patients (20%) with positive core culture, 11 patients (13.75%) had positive surface culture.The commonest organism was E.coli in 13 patients (16.3%) followed by Pseudomonas in 3 patients(3.8%), Klebsiella in 3 patients (3.80%), Staphylococcus aureus in 2 patients (2.5%) & Bacteroides fragilis in 1 patient (1.3%). No salmonella species were isolated. All organisms were tested for sensitivity against quinolones,penicillins, cephalosporins, aminoglycosides & carbapenems.