To investigate the microscopic anatomical features of the associated fascia and mesenteric space during laparoscopic radical gastric cancer surgery, and to provide an anatomical basis for laparoscopic anatomical localization and access. METHODS: The anatomical levels and morphological characteristics of the pancreatic fascia, gastrosplenic ligament and posterior abdominal wall fascial gap formed by the embryonic dorsal gastric tract were observed and described laparoscopically by gross anatomical observation and simulated radical surgery of progressive gastric cancer on a fresh cadaveric specimen. Results: The posterior layer of the dorsal mesentery of the stomach formed the gastrosplenic ligament, the pancreatic fascia, the pancreaticoduodenal fascia, and the anterior lobe of the transverse colonic mesentery as a continuous whole. The posterior border of the anterior renal fascia was the safe operating plane. Conclusions: (1) The extensive perigastric fascia and fascial gap formed during the embryonic period due to intestinal rotation is the key to anatomical localization, tethering and lymph node dissection during laparoscopic radical surgery for gastric cancer; (2) separation along the fascial gap can help improve the safety of laparoscopic operation and the thoroughness of radical treatment; establishing a holistic concept of laparoscopic fascial level anatomy can provide a morphological basis for the design and standardization of laparoscopic surgery. The establishment of a holistic concept of laparoscopic fascial anatomy can provide a morphological basis for the design and specification of laparoscopic surgery.