Background and Aims Laparoscopic anatomical liver resection has become the main surgical approach for space-occupying lesions located in segment Ⅶ (S7) of the posterior lobe of the liver. However, due to the complex anatomical structure of the S7 hepatic pedicle, there may be ischemic lines on the liver surface, but a lack of effective guidance within the liver parenchyma. Given this, our team adopted indocyanine green (ICG) fluorescence-guided puncture positive staining technique for laparoscopic anatomical liver S7 resection. This paper reports on this technique.Methods The clinical data of one patient who underwent laparoscopic anatomical liver S7 resection using the ICG fluorescence-guided puncture positive staining technique in the Department of Hepatobiliary Surgery, Second Affiliated Hospital of Army Medical University were retrospectively analyzed.Results Following successful preoperative 3D reconstruction simulation to obtain the ICG fluorescent staining region of liver S7, an anatomical liver S7 resection was performed using a caudal approach. During surgery, the liver was transected along the fluorescent boundary and the right hepatic vein, with sequential division of the S7 portal vein (P7) and tributaries of the right hepatic vein from S7. The patient recovered well after surgery, and one month later, an abdominal ultrasound review showed that liver S7 had been resected with no signs of tumor recurrence.Conclusion The ICG fluorescence-guided puncture positive staining technique provides excellent assistance in laparoscopic anatomical liver S7 resection, improving surgical safety and ensuring R₀ resection margins.