Superfused synaptosomes have been utilized in studies of neurotransmitter release during 25 years. This review summarizes the aspects of neurotransmission that have been and could be successfully investigated with this technique. The major aim of the article is to draw attention on the versatility of superfused synaptosomes and to suggest how the system could be exploited in clarifying several aspects of synaptic neurochemistry including neurotransmitter transport, receptor localization, receptor-receptor interactions, functional aspects of multi-sited receptor complexes, receptor heterogeneity and mechanisms of neurotransmitter exocytosis-endocytosis.