Use of an appropriate root canal irrigant is essential during endodontic treatment, due to the complex and unpredictable anatomy of the root canal system and limitations in the mechanical instrumentation techniques used to obtain a clean, bacteria-free canal. Several irrigants, such as sodium hypochlorite, chlorhexidine, hydrogen peroxide, and normal saline, have been proposed as canal system irrigants in endodontic treatment. The widely used endodontic irrigant chlorhexidine is a positively charged lipophilic/hydrophobic molecule that interacts with phospholipids and lipopolysaccharides on the bacterial cell membrane. In endodontics, its mode of antibacterial activity is determined by its concentration (0.2% or 2%). This article reviews findings from available endodontic studies on the antibacterial, antifungal, and antibiofilm activities of chlorhexidine. (J Oral Sci 56, 99-103, 2014)