The density of sperm in the water column during the broadcast spawning events of marine invertebrates is often undetermined or reported in terms of fertilization potential. The density of sperm during such events can be determined by directly counting numbers of spermatozoa using a modification of the acridine orange direct count (AODC) technique for enumerating bacteria. A number of variables in the handling and processing of samples may bias estimates. Sample collection in glass and rapid fixation and filtration are necessary for reproducible estimates. Once filtered, samples are stable for many months, and counts on filters that were poorly stained in the field can be enhanced by additional staining with 4'6-diamidino-2-phynlindole (DAPI). The AODC technique was employed to examine the effects of distance from a male colony on density of sperm for the gorgonian coral Plexaura kuna. In situ sperm densities ranged from 0 to 1,000 sperm/ml. Sperm densities in 18 l aquaria containing 20 cm long P. kuna explants reached 100,000 sperm/ml.