Abstract Rodents are the most commonly used experimental models for studying liver structure and function. Detailed knowledge of the liver lobe structure is essential for such studies, yet the biliary system in rodents has not been comprehensively described. This study utilized intravenous injection of indocyanine green (ICG) combined with ligation of the common bile duct, causing extrahepatic bile ducts to appear emerald green under natural light. We observed that bile duct branch of each lobe frequently drained into the superior common bile duct near the gallbladder. Additionally, no previous studies have successfully ligated the portal vein of the left lateral lobe (LLL) in mice, a critical step for models such as portal vein ligation (PVL) or associating liver partition for staged hepatectomy (ALPPS). Using ICG staining and observing liver vasculature under a 25× microscope, we categorized the portal vein of the LLL into four types. Most mice exhibited a distinct portal vein branch for the LLL and left middle lobe (LML), contrary to observations in rats. By selecting the generally and fully exposed type, surgeons or technicians skilled in micromanipulation can successfully ligate the portal vein of the LLL using our method. On postoperative day 3 (POD 3), the LLL remained fresh and alive without apparent necrosis, and the weight ratio between the future liver remnant (FLR) and body, along with positive Ki67 nuclear staining, was significantly enhanced, indicating that ligating the portal vein of the LLL induces liver regeneration.