INTRODUCTION Urinalysis has always been an important diagnostic tool for (extra)renal disorders. Since the 19th century, microscopic urine sediment analysis has been the gold standard (1). Over the past decade, automation (automated microscopy (iQ200) and flow cytometry) has improved the accuracy and productivity of the process (2). After serum chemistry profiles and complete blood counts, urinalysis is the third major in vitro diagnostic screening test (3). The current range of analyses applicable to urine samples is very broad and the preanalytical requirements are extended and strict. The complex pre-analytic process starts with the need for the test and ends with the specimen processing (4). A well-considered strategy with a focus on the pre-analytical phase is essential to improve the reliability of the laboratory results and to decrease the costs of the health care system. The pre-analytical phase of urine testing can be divided into multiple subphases: need for test, specimen collection, transport to the laboratory, specimen receipt in the laboratory, preparation of sample for testing and transport of samples to the proper section in the laboratory (5), all of which can be a source of error. To reduce these errors, more emphasis needs to be put onto the pre-analytical phase (6). In this review, we discuss the pre-analytical challenges of urinalysis, especially for manual and automated particle analysis and test strip analysis.