This study was done to evaluate the frequency of iatrogenic gall bladder perforation (IGBP) in laparoscopic cholecystectomy and to determine its association with gender, adhesions in right upper quadrant and types of gall bladder. This retrospective descriptive study included 200 patients who underwent laparoscopic cholecystectomy for symptomatic cholelithiasis at Jamal Noor Hospital and Hamdard University Hospital, Karachi from January 2007 to January 2009. Video recording of all 200 laparoscopic cholecystectomies were analyzed for the IGBP. The different factors; sex of the patient, type of gall bladder, presence of adhesions in the right upper quadrant, timing of perforation, site of perforation, cause of perforation and spillage of stones were recorded. Data was entered and analyzed on SPSS 15. Pearson Chi Square test was applied to check the significance of these factors in IGBP where applicable. In this study there were 173 females and 27 male patients. IGBP occurred in 51 patients (25.5%) and among them 40(23.12%) were females and 11(40.74%) males. Statistical analysis failed to prove male gender a significant factor in the IGBP (p=0.051). Spillage of stones occurred in 23 patients (11.5% in total study population). In 32(18.49%) patients with chronic calculus cholecystitis IGBP occured while in other cluster of 27 patients suffering from acute cholecystitis, empyema & mucocele, 19(70.37%) had IGBP. Hence the condition of gall bladder (acute cholecystitis, empyema and mucocele) was proved statistically a significant factor in IGBP (p=0.000). Adhesiolysis in right upper quadrant was required in 109 patients in whom 31 patients (28.44%) had IGBP while in 91 patients in whom no adhesiolysis was required, 20 patients (21.98%) had IGBP. Statistically no significant difference was present regarding this factor (p=0.296). In total of 51 patients of IGBP, fundus of gall bladder was the commonest site of perforation in 21(41.18%), followed by body of gall bladder in 18(35.29%) and Hartman's pouch in 12(23.53%) patients. In 27(52.94%) patients, diathermy hook was the cause of perforation followed by grasping forceps in 24(47.06%) patients. In 33(64.71%) patients perforations occurred during dissection of gall bladder from liver bed, in 2(3.92%) during adhesiolysis and in 16(31.37%) during retraction maneuvers. Perforation of gall bladder occurred in 25.5% of patients during laparoscopic cholecystectomy and acutely inflamed and over distended gall bladders were proved significant factor for this intraoperative event.