Glaucoma is a leading cause of irreversible blindness in the world. Intraocular pressure (IOP) plays a key role in glaucoma development and progression. Schlemm’s canal (SC), an important structure of the anterior chamber angle, regulates the flow of aqueous humor and maintains IOP. Because of its special function of aqueous outflow, the SC has been intensive investigated recently. Several characteristics of SC in anatomy, physiology and pathophysiology have been revealed. Compare to normal, glaucomatous SC cells are more sensitive to substrate stiffness, have higher stiffness and and lower porosity leading to higher outflow resistance. And SC collapse caused by acute IOP increase is partially or totally reversal. With advanced inspection techniques, high-quality images of the SC can be obtained in vivo, which facilitates SC quantitative measurements clinically and allows us to investigate a new therapy paradigm for glaucoma. In this review, we summarize the basic and clinical research that focused on mechanisms of aqueous outflow resistance and SC changes in physiological, pathological, and post-treatment states.