This research was initiated to determine the microbial load on endotracheal tube and its elimination by optimizing appropriate dose of gamma radiation. The selected samples were checked for the presence of microbial flora. Different types of media (Nutrient agar, MacConkey agar, potato dextrose agar) were used for enumeration and isolation of microbes. Bacteria were identified by staining and biochemical characterization was done by using API 20E kit. Results of this study showed that Bacillus subtilis, Staphylococcus epidermidis and Staphylococcus aureus were the most common inhabitant of the samples. Neither fungal growth nor gram negative bacteria were isolated. Three doses of gamma radiation (20kGy, 25kGy, 30kGy) were selected for the sterilization treatment of endotracheal tube. The recommended dose for the sterilization of disposable medical devices is 25kGy. However in this study, complete sterility of endotracheal tube was achieved at 30kGy. Thus increased use of gamma radiation for effective sterilization might be helpful in reducing the rate of mortality and morbidity in the Pakistani hospitals, consequently helping to upgrade the public healthcare standard.