To evaluate the effect of preoperative colonoscopic autologous blood labeling to localization for laparoscopic colorectal surgery, data of 30 patients with colorectal neoplasms who underwent colonoscopic autologous blood localization followed by laparoscopic surgery in Shanghai East Hospital, Tongji University from January 2019 to January 2021 were retrospectively analyzed. The location of lesions was determined by local red marks on the serosal surface of the colon during laparoscopic surgery. The clarity of staining and whether there was leakage of injected autologous blood were observed. All patients successfully completed the localization of autologous blood under colonoscopy, without bleeding, perforation, fever or other complications. Laparoscopic exploration showed clear sites of autologous blood staining, without staining dispersion or leakage polluting the operation field. Postoperative pathology confirmed that the margin of the specimen was negative. The injection of autologous blood under colonoscopy is a safe and effective preoperative localization method for laparoscopic colorectal neoplasms surgery, which is worthy of clinical recommendation.