The electroretinogram (ERG) of the isolated bovine retina serves as a proven criterion of retinal activity. It is used as a sensitive pharmacological tool for testing effects of applied drugs and toxins on photoreceptors, and higher order neurons that contribute to the generation of the b-wave. Following isolation and detachment from the underlying pigment epithelium, part of the retina was mounted into a closed chamber and perfused by a nutrient solution. Flow rate of the nutrient solution and its ingredients, incubation temperature and light intensity were optimised empirically to achieve a maximum b-wave amplitude. Under these conditions, a reproducible, high-resolution ERG can be stably recorded for more than 10 h with sufficient oxygenation found to be a prerequisite for the long-lasting stability. Addition of L(+)glutamate to the nutrient solutions was not anymore beneficial for the b-wave amplitude. A well-known inhibitor of oxidative phosphorylation (KCN) and antagonists of voltage-gated Ca2+ channels (isradipine, omega-conotoxin-GVIA and NiCl2) were used to prove the validity of the test system. The recording of the ERG from the isolated and perfused bovine retina serves as a valuable physiological model for a neuronal network in which important questions related to the retinal signalling and metabolism can be investigated.