Purpose: Harvesting at least 12 lymph nodes (LNs) is recommended for adequate tumor staging in colon surgery. Although preoperative endoscopic tattooing has been used for primary localization of tumors, its impact on LN retrieval in colorectal surgery remains controversial. We aimed to investigate the relationship between preoperative tattooing and LN retrieval after laparoscopic rectal resection. Methods: We reviewed the records of 92 patients with rectal cancer who underwent laparoscopic resection from January 1, 2018 to December 31, 2019. Patients were categorized into 2 groups according to whether preoperative endoscopic tattooing was performed. The rate of adequate LN retrieval (≥12) was compared. Results: The tattooed and non-tattooed groups comprised 49 and 43 patients, respectively. In the tattooed and non-tattooed groups, the rates of adequate LN retrieval were 75.5% and 55.8%, respectively (P=0.046). Univariate analysis revealed that female sex, tattooing, LN metastasis status, pathological pathological stage (p-stage), and LN dissection were predictive factors for adequate LN retrieval. In the multivariate analysis, female sex (odds ratio [OR], 3.34; 95% confidence interval [CI], 1.15–9.73; P=0.027), tattooing (OR, 2.87; 95% CI, 1.03–7.94; P=0.043), and p-stage (OR, 3.34; 95% CI, 1.04–10.75; P=0.043) were independent predictive factors for adequate LN retrieval after surgery. Conclusion This study revealed that preoperative endoscopic tattooing was statistically significantly associated with adequate LN retrieval in patients with rectal cancer who underwent laparoscopic rectal resection. Preoperative endoscopic tattooing should be considered to improve disease assessment and avoid stage migration.