Background: Intraoperative localization of polypectomy sites is critical for laparoscopic and open surgery, but conventional methods of marking the colorectum are sometimes unreliable. A new two-step method for marking polypectomy sites for identification during laparoscopic and open operations is described. Methods: Eighteen patients with postpolypectomy lesions necessitating preoperative marking were enrolled in this study. A physiologic saline solution was endoscopically injected into the submucosa to produce an artificial submucosal elevation (pseudopolyp). A small volume of India ink was then injected into the submucosal elevation (pseudopolyp) with a separate needle. Results: The two-step method was easily applied for all lesions without complication. At surgery, all lesions were immediately visualized. Conclusion: The two-step tattooing method proved to be easy, safe, and accurate for marking polypectomy sites.