The common reason of pathological high intraocular pressure in glaucoma is the obstruction of aqueous humor outflow, so the morphological observation and functional study of aqueous humor outflow channel is the focus of glaucoma prevention and treatment. The study of the morphology and function of aqueous humor outflow channel is of great significance for the study of the pathogenesis of glaucoma, the selection of surgical methods and the development of targeted drugs. The imaging methods of aqueous humor outflow include anterior segment optical coherence tomography (AS-OCT), OCT angiography (OCTA), ultrasound biomicroscopy (UBM) in vivo imaging, static and real-time aqueous humor angiography, etc. OCT and UBM can evaluate the structure of aqueous humor outflow channel, while aqueous humor angiography is a functional assessment. At present, there is no precise relationship between the outflow characteristics of aqueous humor structure and the value of functional flow. Aqueous humor angiography can be used to study and evaluate the drug efficacy of glaucoma. It is also helpful to guide trabecular meshwork targeted minimally invasive glaucoma surgery to achieve a better effect of lowering intraocular pressure. However, aqueous humor angiography is invasive and can only be used in the operating room. In the future, the method of evaluating aqueous humor outflow pathway needs to be further improved in order to better understand the structure of aqueous humor outflow pathway and anterior segment structure and improve the level of diagnosis and treatment of glaucoma through the combination of structural and functional evaluation. (Int Rev Ophthalmol, 2020, 45: 397-403)