Fig. 12

Individual unprocessed 10 × magnification micrographs of histological Masson’s Trichrome-stained top, center, and bottom layer of male rat’s distal left-side BEFAF segment before (left panel) and after (right panel) manual image stitching procedure to generate one seamless image. The digital stitching process starts with opening all three raw unprocessed images in Adobe Photoshop and extending the canvas of the first image down. Then a second image was dragged as a layer onto the first image trying to align and eliminate overlapping features (indicated by yellow arrows). Lastly, the third image was placed underneath. The exposure of each layer was adjusted to match the previous layer using “Image > Adjustments > Levels” tool by sliding the input levels to the left or to the right. The darker edges of each image (if any) were eliminated by applying “Dodge” tool. Finally, the layers were merged and cropped to eliminate blank spaces on the left and/or right sides. The final seamless image was adjusted for contrast, exposure, and sharpness. Scale bar = 200 µm