The detection of fungal elements and their characterization in patient specimens provides fundamental information. Culture-based methods, though often slow, may yield the specific etiological agent, and may allow susceptibility testing to be performed. Proper collection and transportation of the specimen is essential. Particularly, sterile materials are important for diagnosis of invasive fungal infections. Therefore, culture and direct microscopy should be performed on all suitable clinical specimens when fungal disease is suspected. Numerous different media for culturing and identifying fungi are available, and those important for diagnosing mycoses as well as the most important staining methods for direct microscopy are described.