FIG. 3.

Two-photon live animal imaging of skeletal reporters in pristine bone. (a) Two-photon z-stack through mouse calvarial bone carrying a Col2.3emd reporter (green) and Alizarin mineral label (red). Red square on leftmost diagram indicates the x-y imaging location. Ten micrometer slice shows collagen fibers, white arrow. Thirty-four micrometer slice contains osteocytes, red arrow. (b) Parietal bone carrying a Col3.6topaz reporter (green) and rhodamine-labeled dextran in the vasculature (red). Osteoblasts localize in the area between the vasculature and the endosteal surface of bone. (c) Stereomicroscopy (leftmost panel) and two-photon z-stack of suture line containing SMAAemd osteoprogenitors (green) and rhodamine- labeled blood vessels (red). (d) z-Stack of SMAAemd osteoprogenitors shown in inset of (c). (e) Stereomicroscopy (leftmost panel) and two-photon z-stack of large blood vessels (*) in the parietal bone labeled with SMAAemd. Asterisks in (e) indicate SMAA⁺ blood vessels. (f) Smooth muscle cells labeled with SMAAemd wrap around rhodamine-labeled blood vessels. Color images available online at www.liebertpub.com/tec