Objective To investigate the technique and dosage selection of indocyanine green(ICG) fluorescence staining in laparoscopic anatomical hemihepatectomy. Methods A retrospective cross-sectional study was conducted. The clinical date of the patients who underwent laparoscopic anatomical hemihepatectomy in the Cancer Hospital of the Chinese Academy of Medical Sciences from October 2020 to October 2023 was collected and analyzed, and the management of the Glissonean pedicle, the method and effect of ICG fluorescence staining during the operation, the dose of ICG injection, and the postoperative recovery were analyzed. Results A total of 91 laparoscopic anatomical hemihepatectomies were enrolled in this study, including 28 right hemihepatectomies and 63 left hemihepatectomies. The Glissonean pedicle was dissected intra-sheath in 9 cases and extra-sheath in 82 cases. ICG fluorescence staining was all performed using the negative staining method, of which 69 cases(75.8%) were successfully stained. The success rate of staining in the extra-sheath dissection and low-dose ICG group was higher than that in the intra-sheath dissection and high-dose ICG group. The average operation time was (168.5±32.2) minutes, the intraoperative bleeding volume was (152.4±56.3) ml, and the intraoperative blood transfusion rate was 6.6% (6/91), the average postoperative hospital stay was (8.5±2.6) days. One case was converted to laparotomy due to exophytic growth of the tumor compressing the Glissonean pedicle. Four cases had Clavien-Dindo Ⅰ-Ⅱ complications, all of which improved after treatment. There were 3 cases of grade Ⅲa complications, all of which were caused by bile leakage and abdominal cavity infection. They were cured by puncture and drainage. And there were no serious complications above grade Ⅲb. Conclusions In laparoscopic anatomical hemihepatectomy, the ICG fluorescence staining method was recommended to use the negative staining method of the extra-sheath dissection of the Glissonean pedicle, and a lower dose of ICG could help to increase the success rate of fluorescence staining.