Light-microscopic demonstration of methylene blue accumulation sites in mouse brain after supravital staining.

Mouse brains were stained supravitally with methylene blue and studied in paraffin sections by light microscopy. In the perikarya, the dye was found to bind primarily to the nuclei; only slight staining of the cytoplasm was observed. Dye accumulations within nerve fibers were found in the nodes of Ranvier and in the varicosities of the unmyelinated endings. Specific dye binding in dendrites corresponded mainly to beads and spines. The accumulation sites in terminal neuronal processes appeared to be closely related to the plasma membrane. These morphological data would explain the neurophysiologically proven interaction of the dye with calcium-binding sites in membranes.