The potential fertility of a semen sample will probably depend on that a sufficient number of viable, morphologically normal and functionally competent spermatozoa reach the oviducts and establish an oviductal reservoir, accomplish normal fertilization, and contribute to sustain embryo development. In vitro sperm evaluation, to have any predictive value of its fertility potential in vivo, should include the assessment of as many morphological and functional sperm characteristics as possible, especially for the evaluation of frozen-thawed spermatozoa and of new methods for semen cryopreservation. The new computerized systems for sperm motility and morphometry analysis, and fluorescence technologies, enable multiple and objective measurements of structural and functional sperm parameters to be adquired in a short time. In addition, assays based on in vitro fertilization provide valuable information about the functional ability of spermatozoa when interacting with the oocytes. In the present work some of the in vitro methods currently used for bovine sperm evaluation are reviewed, though not exhaustively, covering some attributes of the sperm cell that are useful in estimating its functional ability.