Infection is a major impairment in delayed and nonhealing chronic wounds. Cultures of chronic wounds are not routinely performed unless there is a clinical suspicion of infection. The culture is then indicated to identify the causative organism and to guide antibiotic therapy, systematically and topically. The deep tissue biopsy is regarded as the gold standard for identification of wound bioburden and clinical infection. However, controversy persists among practitioners as to the relevancy of doing cultures and the technique to use. This article will explore the various techniques used in clinical practice to obtain cultures for diagnosing wound infections.