Considerable morphological variations are observed in the ossicles among lipotyphlans. 3D analysis was conducted to compare the middle ear among lipotyphlans to clarify how the morphological traits of the ossicles have changed during the adaptational evolution, since lipotyphlans have adapted to various lifestyles, such as terrestrial, semi-subterranean and subterranean. The results of the PCA and CVA indicated that subterranean species possess the morphological characteristics such as less development orbicular apophysis and shorter AP. Hypertrophied orbicular apophysis and longer AP were confirmed in the terrestrial and semi-subterranean species. The composite phylogenetic tree and PCA data pointed out that the ossicles of erinacidae, soricidae, crosidurinae and talpidae were distinguishable. However distribution area of erinacidae was confirmed near that of crocidurinae in the PCA chart. The plots of semi-subterranean and aquatic/subterranean moles in talpidae were closely distributed to those of soricinae. Although the middle ear structure in lipotyphlans is partially determined by phylogenetic factor, the morphological characteristics adaptational to each lifestyle also can be observed. From the present results, it is suggested that the middle ear structure may be functional-morphologically supported by the lifestyle strategy in lipotyphlans.