The ovipositor of striped bitterling Acheilognathus yamatsutae was subjected to ultrastructure and histochemical analysis during spawning season using light and electron microscopy. Although the ovipositor of A. yamatsutae is a long cylindrical tube with smooth external surface, it was possible to confirm the presence of well-developed fingerprint structure using scanning electron microscopy. Internal aspect analysis of ovipositor revealed formation of 5-8 longitudinal folds. Cross section analysis revealed that the ovipositor is composed of an outer epithelial layer, a mid connective tissue layer, and an inner epithelial layer. The outer epithelial layer contains 7-9 cell layers composed mainly of epithelial and mucous cells. Result of AB-PAS (pH 2.5) and AF-AB reaction showed that mucous cells contained mainly acidic carboxylated mucosubstances. The connective tissue layer was loose and made mainly of collagen fibers and some muscle fibers, along with blood vessels and a small number of chromatophores. The inner epithelial layer, which is a single layer, is composed of columnar epithelia. Observation under transmission electron microscope enabled distinction of the outer epithelial layer into superficial, intermediate and basal layers. Although the types of cells in the superficial tissue layer were diverse, they all shared the development of glycocalyx covered microridges. The majority of epithelial cells in the intermediate layer were cuboidal shaped, while those in the basal layer were columnar. Two types (A and B) of secretory cells were observed in the outer epithelial layer. The connective tissue layer had two types of chromatophores including xantophore and melanophore, in addition to a well-developed nerve fiber bundles. Columnar epithelial cells, mitochondria-rich cells and rodlet cells were observed in the inner epithelial layer. Microvilli were well developed on the free surface of columnar epithelial cells.