Figure 3

Micrographs of the exposed deep fascia without hook-holding used in plunge freezing followed by freeze-substitution technique. (a) Hematoxylin–eosin (HE) stain. The deep fascia has abundant vessels, particularly, the first superficial layer (*) with collagen fibers extending in various directions. The second intermediate layer was formed by strongly wavy and thick bundles of collagen fibers (**). The third, deepest layer displayed straight and thin bundles of collagen fibers (#). Scale bar = 20 µm. (b) HE stain. The nerve tissue was identified (arrow) in the first superficial layer. Scale bar = 15 µm. (c) Elastica Van Gieson (EVG) stain. Collagen bands were forming clearly different layers. Elastic fibers (arrow) were identified in all layers of the deep fascia. Scale bar = 20 µm.