Fig. 11
Individual unprocessed 4 × magnification micrographs of histological Masson’s Trichrome-stained sections of male rat’s middle left-side BEFAF segment before (upper panel) and after (bottom panel) manual image stitching procedure to generate one seamless image. This flap with an estimated 100% viability was harvested 48 h post-reconstructive surgery, where the left-side flap was exposed to 4 h PI, 2 h REP1 and 4 h of SI (AVO), whereas right-side flap was exposed to 4 h PI, 6 h REP1 and 4 h of SI (AVO). The digital stitching procedure started with opening all three raw unprocessed images in Adobe Photoshop and extending the canvas of the first image to the right side. Then a second image was dragged as a layer onto the first image trying to align and eliminate overlapping features. Lastly, the third and fourth images were placed underneath. The exposure of each layer was adjusted to match the previous layer using “Levels” properties. The darker edges were eliminated by using “Dodge” tool. Finally, the layers were merged and cropped to eliminate blank spaces on the top and bottom sides. The background of the final image was cleaned from debris by selecting the background with the “Quick Selection” tool, deleting it, and replacing it with the white color. The final image was then adjusted for contrast and sharpness. Scale bar = 1 mm