Figure 4. Confocal microscopy of laser-induced choroidal neovascularization (CNV), showing extent of endothelial cell proliferation in control animals. A, B: In this flatmount preparation of a control choroid at 7 days post treatment, the retina has been removed to expose a typical laser lesion through Bruch’s membrane and into the choriocapillaris. A: Maximum projection (brightest voxel displayed for each Z-column) images of immunolocalization for cytoskeleton (f-actin, green), nuclei (Heochst, blue), and endothelial cells (isolectin B4, red) are shown on the left, along with a merged image of the three labels. The initial lesion (50 µm diameter) is apparent in the f-actin image, while endothelial cells (capillaries) are seen to fill the site as proliferation proceeds. B: The same lesion as in A, presented as an orthogonal cut view. The large square image represents a single layer in a confocal stack of images, as viewed above from the retina, with the three labels merged. Lateral views of this image stack are shown at the top and to the right; the blue line in these views indicates the level within the stack of the view shown in the square. The lateral view at the top is from the position of the green line in the large square; the lateral view to the right is from the position of the red line in the large square. C, D: Images are organized the same as in A and B and depict a typical laser lesion of a control retina at 14 days post treatment. White circles represent the 50-µm-diameter initial laser lesion. Magnification bars equal 50 µm.