Introduction: Natural orifice specimen extraction surgery (NOSES) has been widely regarded as a new technology in minimally invasive surgery. Meanwhile, carbon nanoparticles have been increasingly used for lymph node tracing in colorectal cancer surgery.Aim: To evaluate the effectiveness and feasibility of carbon nanoparticle-assisted natural orifice specimen extraction surgery with left colic artery preservation for total laparoscopic colorectal resection. Material and methods: We retrospectively reviewed the medical records of 83 patients diagnosed with sigmoid colon cancer or mid-and upper-rectal cancer from October 2017 to June 2020. These patients were divided into the NOSES group who underwent left colic artery preservation NOSES, being injected with a carbon nanoparticle suspension under colonoscopy the day before surgery, and the LA group, who underwent left colic artery preservation laparoscopic surgery. Surgical outcomes were retrospectively analyzed. Results: The mean number of harvested lymph nodes (p < 0.001) in the NOSES group was higher than in the LA group. Conversely, as regards pain score (p < 0.001) and postoperative hospital stay (p = 0.035), the LA group has higher mean values. The incidence of perioperative complications (p = 0.385) was 5.3% for the NOSES group compared to 13.3% for the LA group. Conclusions: Preoperative colonoscopic injection of a carbon nanoparticle suspension is a feasible and practical solution to dissect lymph nodes surrounding the inferior mesenteric artery without affecting the left colic artery in patients with colorectal cancer and about to receive NOSES. Moreover, NOSES combined with this approach leads to less postoperative pain and shorter hospital stays.