FIG. 2.

Osteoblasts, osteocytes, and mineralization in the mouse calvarium. (a) View of the left parietal lobe of a calvarium from a Col2.3emd mouse showing osteoblasts (green) and mineral surface labeled with Alizarin complexone (red). The green, red, and brightfield channels are overlaid to show the colocalization of the osteoblasts and new bone label. The sagittal and coronal suture lines also contain osteoblasts, indicated by the black asterisks (*). The horizontal line indicates the approximate location of the cross-sectional view shown in (d). The anatomical locations lambda and bregma are labeled λ and br., respectively. (b) Inset from (a) shows a magnified view of the marrow spaces located in the parietal bone. Blood vessels observed with brightfield optics (white arrow in c) localize to the inside of the marrow spaces (white arrow in b, same location as c). (c) Brightfield channel of (b) showing the organization of vasculature (white arrow). (d) Histological section of (a) showing Col2.3emd osteoblasts and osteocytes (white arrow) in the marrow spaces overlaid with a differential interference contrast channel showing cortical bone. Mineralization label (red) is located on the surface of endosteal marrow spaces. The white line indicates the 139-μm total depth of the two-photon z scan shown in Figure 3a. (e) Emerald signal of (d) showing osteocytes (white arrow) in the cortical phase. Color images available online at www.liebertpub.com/tec