The purpose of this study was to present the evolution of ideas on the examination of urine from antiquity till our days. A thorough study of texts, medical books from antiquity till twentieth century along with a thorough review of the available literature in PubMed was conducted. The first observation on urine examination can be traced back to the Babylonian and Sumerian texts. Almost all physicians in antiquity including Hippocrates referred to the value of urine examination in the diagnosis and prognosis of diseases. The construction of first compound microscope lead to the examination of urine sediment and the development of Urine Cytology which was revolutionized during the twentieth century with the studies of important cytologists such as George Papanicolaou, Geoffrey Krabbe, and Leopold Koss. The introduction of molecular tests in the diagnosis of urothelial cancer inaugurated a new era in the study of urine cytology. The history of urine examination spans a period of 6,000 years. The application of microscope in the examination of urine sediment during the nineteenth century established urine analysis as an important diagnostic tool in clinical practice. Diagn. Cytopathol. 2015;43:1020–1036. © 2015 Wiley Periodicals, Inc.