The chapter gives a brief overview of surgical asepsis and antiseptic measures. Surgical asepsis is defined as prevention of access of microorganisms to a surgical wound. Antiseptic measures are the measures used to destroy bacteria or remove them from all objects coming in contact with the wounds. Surgical wound sepsis can be prevented or minimized by following operation theater (OT) discipline, and proper instrument sterilization. The World Health Organization checklist (regarding patient details, type of surgery, site of operation, etc.) is a universal tool to improve patient safety and should be completed for every patient coming to theater. Protocol for waste disposal is also given in an illustrated manner. Sterilization is the process of killing all microorganisms including bacteria, virus, fungi, parasites and spores. Disinfection is the process of killing all microorganisms except spores. Methods of sterilization are also stated like boiling, autoclaving, chemical methods, dry heat, ethylene oxide, formaldehyde, and gamma radiation.