Male subfertility is a common problem with a complex etiology, requiring a complete andrological work-up for proper diagnosis. The male reproductive tract is controlled by a well-balanced hormonal system, in which hypothalamic (GnRH), pituitary (LH, FSH) and testicular hormones (androgens, inhibin B) participate. Any disturbance of this hormonal system may therefore lead to testicular dysfunction and interfere with the spermatogenesis process. In addition, also other components along the ductal system, such as epididymis, prostate and seminal vesicles, that improve sperm fertility by contributing their secretions to the semen, might function inadequately and thus fail to enhance the fertilizing capacity of the sperm cells. External factors (heat, chemicals, life style) and anatomical abnormalities (varicocele) were shown to have a negative influence on male fertility. In a number of patients genetic defects can be identified as the cause of their infertility. Laboratory tests are available to assess hormone concentrations, semen composition, accessory gland function and sperm cell function. Conventional semen analysis includes the determination of sperm concentration, semen volume, sperm motility (qualitative and quantitative), sperm morphology, sperm cell vitality, pH, leucocytes and antibodies. The usefulness of the determination of these parameters as predictor of fertility appears to be rather limited, however. Therefore, alternative tests, some based on more functional aspects (sperm penetration, capacitation, acrosome reaction), have been developed. Furthermore, there is an increasing attention for the assessment of DNA integrity, for instance by the flowcytometer-based Sperm Chromation Structure Assay (SCSA), as an additional or alternative parameter of sperm quality. It is likely and desirable that further assays with better predictive value are being developed in the near future.