Objective: To evaluate the clinical efficacy and safety of microscopic radiofrequency ablation in treating of distichiasis and aberrant lashes. Methods: We performed a retrospective analysis on 18 patients with trichiasis (distichiasis and aberrant lashes) who underwent microscopic radiofrequency ablation treatment. All patients were followed up for a minimum of 3 months after the operation. The outcome measures included trichiasis clearance rate, the number of treatment sessions needed to achieve success, complications, and patient satisfaction. Spearman rank correlation analysis was used to evaluate the correlation between the baseline characteristics of patients and the number of treatment sessions. Results: The study included 18 patients with a total of 30 eyes. Immediately after the operation, the trichiasis clearance rate achieved through microscopic radiofrequency ablation was 100%. Among the eyes, 5 eyes (16.7%) were successful treatment after just one session, 11 eyes (36.7%) required 2 treatment sessions, 13 eyes (43.3%) needed 3 treatment sessions, and 1 eye (3.3%) required more than 3 treatment sessions to achieve success. The major complications included dry eye in 13 cases (43.3%) and eyelid margin inflammation in 1 case (3.3%). Notably, no severe complications such as eyelid deformity or scarring were observed. According to the patient satisfaction survey, 16 cases (88.9%) reported being very satisfied, and 2 cases (11.1%) were basically satisfied, resulting in an overall satisfaction rate of 100%. The correlation analysis indicated that the number of trichiatic eyelashes was significantly positively correlated with the number of treatment sessions (rs=0.672, P=0.002). Conclusions: Microscopic radiofrequency ablation for distichiasis and aberrant lashes treatment boasts serveral advantages, including precise localization, effective destruction of hair follicle, short treatment duration, minimal trauma, rapid recovery, and high safety. With a high level of patient satisfaction, it serves as an effective treatment method for distichiasis and aberrant lashes.