Neurons transmit information by exocytosis of synaptic vesicles (SV), which contain neurotransmitter. Exocytosis is followed by efficient retrieval of the plasma membrane by endocytosis to generate a new SV. SV retrieval supports multiple cycles of synaptic transmission. Over the years, styryl dyes have been widely used to probe the mechanism of SV recycling in the processes of cultured neurons. The styryl dye method is complementary to electrophysiological measurements or genetic reporter approaches. Owing to their ease to culture, cerebellar granule neurons provide a robust neuronal model system for the assay. These cells are readily transfected with various DNA constructs to study the function of exocytic or endocytic proteins in SV recycling.