A good understanding of anorectal physiology is essential for the diagnosis and appropriate treatment of various anorectal disorders, such as fecal incontinence, constipation, and pain. This article reviews the physiology of the anorectum and details the various investigations used to diagnose anorectal physiology disorders. These anatomic and functional tests include anal manometry, endoanal ultrasound, defecography, balloon expulsion test, magnetic resonance imaging, pudendal nerve terminal motor latency, electromyography, and colonic transit studies. Indications for investigations, steps in performing the tests, and interpretation of results are discussed.