A combined light and electron microscopic method for the visualization of the same in vitro neuron by radioautography and serial sections

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Abstract
A technical improvement is described which makes it possible to study, at the ultrastructural level, a dopaminergic neuron which was previously identified by light microscopy. Primary cultures of virtually pure mesencephalic neurons from mouse embryos were obtained. These cultures were kept for 6 days, then incubated with [3H]dopamine fixed and embedded in Epon. The dopaminergic neurons were 1st visualized by radioautography directly through Epon blocks in toto by light microscopy. In a 2nd step, ultrathin sections of the identified dopaminergic cells were prepared and the neurons observed at the EM level. The dopaminergic nature of these neurons was regularly checked by radioautographic control on some selected ultrathin sections.