The Contribution of Dithiothreitol Separation to the Morphological Studies of the Epidermal and the Epithelial Basement Membranes :

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There are several methods of epidermal-dermal separation. In many cases the separation occurs at the lamina lucida, and the cellular surface of the lamina densa is exposed. The dithiothreitol separation is the only method to induce epidermal-dermal separation below the lamina densa, resulting in the exposure of the interstitial surface of the lamina densa. After this separation, the morphology of the anchoring fibrils is clearly observed, and immunohistochemical and the lectinhistochemical studies are possible on the interstitial surface, using colloidal gold-or ferritin-conjugated antibodies or lectins. SEM observations on the interstitial surface of the lamina densa and the upper surface of the lamina fibroreticularis are also possible with this separation method. The dense meshwork structure of the lamina densa, the anchoring fibrils and the aparse meshwork with thin collagen fibrils in the lamina fibroreticularis were three-dimensionally observed. Colloidal gold particles, which were difficult to identify in the secondary electron images, were observed due to their stark contrast against the background in the backscattered electron images. The distribution of the colloidal gold-conjugasted antibody was three-dimensionally observed by backscattered electron imaging.

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  • Connective tissue

    Connective tissue 31 (3), 181-187, 1999-09-25

    日本結合組織学会

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