A 14-year-old, spayed female Cocker Spaniel was presented to the Boren Veterinary Medical Teaching Hospital at Oklahoma State University with acute loss of vision in the right eye and a history of intermittent bloody diarrhea of unknown duration. Small, white, plaque-like lesions in the retina and subretina were visualized by direct ophthalmic examination, and aspirated with ultrasound-guidance. A direct smear of the subretinal fluid was highly cellular and contained large numbers of pleomorphic organisms consistent with Prototheca sp. The structures were round, oval, or elongated, 4-6 microm width and 8-16 microm in length and surrounded by a thin, clear cell wall. Small, central, pink to purple nuclei were observed in some organisms, but in most, the nuclei were obscured by a deeply basophilic, granular cytoplasm. Some organisms contained endospores. Negatively-stained structures of similar size and shape were considered to be empty casings (theca) of ruptured sporulating and nonsporulating forms of the organism. Protothecosis usually is a disseminated, fatal disease in dogs. The Prototheca organisms observed in this case showed characteristic morphology, illustrating the ability to diagnose protothecosis in cytologic samples.