Computer-assisted combined indocyanine green (ICG) molecular fluorescence imaging technology can be used for preoperative planning and intraoperative detection from three-dimensional (3D) morphological anatomy and level of cellular function to guide the anatomical, functional and radical hepatectomy of liver tumor. This technology has received wide acceptance and has shown important diagnostic and therapeutic value. This guideline is intended to standardize the application of computer-assisted combined ICG molecular fluorescence imaging for accurate diagnosis and treatment of liver tumors in the following aspects: (1) the workflow of 3D visualization technology; (2) the mechanism and application flow of ICG molecular fluorescence imaging; (3) clinical application of 3D visualization technology and virtual reality technology; and (4) clinical application of ICG molecular fluorescence imaging. ICG molecular fluorescence imaging can help to define tumor boundary, determine hepatic segment and hepatic lobectomy tangent at the molecular and cellular level, and detect small lesions or metastases. According to the fluorescence signal characteristics of liver tumors and combined with rapid frozen pathological examination during operation, the differentiation degree of liver space-occupying lesions (such as primary liver cancer) can be preliminarily determined, and residual tumors and biliary leakage on the hepatic section can be detected after hepatectomy. Computer-assisted ICG molecular fluorescence imaging in the diagnosis and surgical navigation of liver tumors provides a new approach to digital diagnosis and treatment of liver tumors. With its development in clinical practice and the technological innovation, this technology will be further improved to allow more accurate diagnosis and treatment of liver tumors.