Confocal Laser Scanning Microscopy of Small-Intestinal Mucosa in Celiac Disease

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Abstract
Background: Light microscopy of jejunal biopsy specimens is routinely used to diagnose celiac disease. Confocal laser scanning microscopy offers research advantages, since thin optical sectioning can be performed without mechanical damage to the specimen. Fixed as well as non-fixed specimens can be studied. Methods: Confocal laser scanning microscopy was used to study the staining of wheat germ agglutinin, which binds to glycoconjugates of the enterocyte, and rhodamine phalloidin, which binds to the F-actin of the cells. Results: In healthy mucosa the wheat germ agglutinin labeling showed a strong and punctate staining of microvilli, which outlined a convoluted surface. Phalloidin-labeled actin formed a three-dimensional cage at the cell membrane. In the crypt-hyperplastic mucosa, both staining patterns were irregular, and the cytoskeleton was disorganized. Conclusions: Confocal laser scanning microscopy offers the possibility to study the distribution of surface and cytoskeleton markers in thick, structurally intact specimens.