Whole body of a Korean male cadaver was serially sectioned to make anatomical images. Structures in the anatomical images should be segmented to make three-dimensional images of the structures. Purpose of this research is to prepare the segmented images of lower limb in detail, which are distributed to help other investigators make 3D images and virtual dissection software of lower limb. One-hundred fourteen structures (a skin, 32 bones, 7 knee joint structures, 60 muscles, 7 arteries, 7 nerves) of left lower limb were decided to segment in 976 temporary segmented images (PSD file, intervals 1.0 mm, resolution 1,200×1,100) including anatomical images. On the Adobe Photoshop, selections which fit the structures’ contours were drawn automatically, semiautomatically, or manually; subsequently, the selections were put into the layers. After filling the selections with colors, the temporary segmented images were converted to 996 segmented images (TIFF files). The segmented images were staked to make coronal and sagittal segmented images for verifying segmentation. The corresponding segmented and anatomical images can be used to make three-dimensional images of the 114 structures and virtual dissection software, which are helpful in anatomy study of lower limb. The segmentation techniques of this research can be used to segment many structures in other images quickly and correctly.