In phase contrast and scanning electron microscopy, diverse structural forms of cells tenaciously adherent to glass coverslips were observed in the culture of the corpus callosum and cavum septum pellucidum from postnatal rats. In day 1 culture, many of the cultured cells were round, with well spread peripheral cytoplasm which appeared homogeneous. Cell organelles aggregated mainly around the reniform or round nucleus. Some cells showed spinous projections. In day 3-5 culture, the cells became irregular, sending out long branching pseudopodial processes; often they displayed a vacuolated cytoplasm. The cultured cells were highly phagocytic, as shown by their uptake of colloidal carbon particles and latex beads, in light microscopy and scanning electron microscopy, respectively. Cytochemical studies have shown that the cells were peroxidase-negative but were strongly positive for non-specific esterase, similar to the amoeboid microglial cells in the postnatal corpus callosum. On the basis of their structural features, both in phase contrast and scanning electron microscopy, experimental as well as cytochemical properties, it is concluded that the cells in the present culture are in fact amoeboid microglial cells which are active macrophages in the developing corpus callosum.