Fig. 4.

3D imaging of the vasculature obtained with SPIM. A, Maximum intensity projection of an anti-CD31 labeled mouse heart apex. SPIM-excitation laser, 635nm; emission filter, 670 nm. B, 3D-visualization the same data shown in A. C.1, Single SPIM image of a FITC-lectin labeled mouse heart. The low contrast between background (BG) and labeled vasculature makes it difficult to distinguish between them. Autofluorecence and stripes artifact harm the image quality. C.2, SPIM-image of a lectin-647 labeled mouse heart. The BG is minimized significantly by the absence of autofluorecence in the far-red channel. The stripes artifact almost entirely disappear. D, Single image of a SPIM image stack in which the macro- and micro-vasculature were both labeled with FITC-lectin (white). E, 3D volume rendering of the stained vasculature, stack size: 1945x1214x380 mm, visualized with VolView3 software (Bangham laboratory). F, Zoom of the indicated zone in E. All scale bars in all images, 100µm.