Several selection techniques are available for processing spermatozoa. Apart from sperm washing to remove seminal plasma, only “swim-up” and colloid centrifugation have been used to any extent to prepare spermatozoa for in vitro fertilization, and only colloid centrifugation has been used to prepare sperm samples for artificial insemination. Single-layer centrifugation (SLC) through a species-specific colloid has been shown to be effective in selecting spermatozoa with good motility, normal morphology and intact chromatin in a range of species. This method is less timeconsuming than swim-up, and has been scaled-up to allow whole ejaculates to be processed in a practical manner. The applications of SLC are as follows: to improve sperm quality in insemination doses or in samples for in vitro fertilization, to increase the shelf life of normal sperm doses, to remove pathogens (viruses, bacteria), to improve cryosurvival by removing dead and dying spermatozoa before freezing or after thawing, to select spermatozoa for intracytoplasmic sperm injection, and to aid conservation breeding.