Carotenoid-rich oil globules were isolated from the cytoplasm of the binucleate dinoflagellate, Perinium foliaceum. These orange globules were collected from ruptured ceUls by ultracentrifugation on a sucrose density gradient, and checked for purity by electron microscopy. The osmiophilic globules were assayed for lipid (including pigment) (5), who discovered that the carotenoid globules are highly organized in a flattened triple-membrane-enclosed sac, designated an "eyespot." This organelle is composed of two layers of osmiophilic globules (80-200 nm in diameter) separated by some granular material in the center. The eyespot has a trapezoid shape, about 6 Am by 3 ,um and 350 nm thick, and is always located ventrally at the anterior end of the sulcus (5).P. foliaceum is a binucleate dinoflagellate, and the presence of a eucaryotic nucleus, in addition to the typical dinocaryotic one, logically has been attributed to the existence of a chrysophyte-like endosymbiont which is inside of the heterotrophic dinoflagellate (4,16,18). The nucleus, chloroplasts, and other organelles of the symbiont are surrounded by a unit membrane (8). The dinocaryotic nucleus and other organelles, including the eyespot, pertain to the heterotrophic dinoflagellate, not its endosymbiont (18). Red-orange lipid globules have been observed in other heterotrophic dinoflagellates but these "colored oil droplets" have never been isolated for biochemical study.Dodge and Crawford (5) To eliminate the possibility of bacterial contamination, axenic cultures were also prepared using a synergistic penicillin-streptomycin mixture (4 and 0.8 mg/ml) with no antagonistic chloramphenicol (see 14). The cells were cultured in Fernbach flasks (1.5 liters) in GPM medium (10).