Figure 7

Imaging abnormal features of developing mammary glands using optimal SHG-B/Carmine Alum/SHG-F approach. A. A three color, single slice (XY) from a Z-stack of images of 46 μm thickness is presented from a Carmine Alum whole mount (HAI-1 mouse gland). B. The three-color, 3D projection is shown (3D) as well as orthogonal views which were acquired separately for just the SHG-B and SHG-F signals using a narrower bandwidth in the ChS (META) detector for SHG-B. Arrows in the lower magnification upper panel indicate differences between the XY and 3D views of the lateral branches, which otherwise look similar indicating that they are mainly viewed in a single XY orientation relative to the stage. C. A higher magnification view of the boxed inset region in A (XY) reveals details of Carmine Alum staining and SHG-B and SHG-F. The fibers visualized using SHG are concentrated near the duct and lateral buds (asterisks in A, B and C). The dotted arrows shown in C indicate fibers (SHG-F) between and around the lateral buds. D. A higher magnification view of the boxed inset region in B reveals the 3D view of Carmine Alum staining with SHG-B and SHG-F. E. Orthogonal views including XY, XZ, and YZ reveal two lateral buds viewed as two color image of SHG-B and SHG-F (E). F. A conventional bright field image of a similar duct taken using a Nikon upright microscope equipped with a 10X/ 0.3 N. A. Nikon lens. Scale bars = 50 μm in A (top panel) and 20 μm elsewhere except F, 100 μm.