Abstract
Fixation of tissues for postembedding gold-lectin histochemistry calls for a fixative that preserves the ultrastructure of the tissue, has no inhibiting effect on lectin binding sites, and binds the glycoconjugates, which carry these lectin binding sites within the tissue so strongly that they cannot be washed out during the dehydration and ensuing embedding steps. There is no “one and only” fixative that has all these properties, and every fixative used for postembedding gold-lectin histochemistry must be viewed, at best, as a compromise. One common fixative, more or less exhibiting the properties required, is a mixture of 4% formaldehyde and 0.5% glutaraldehyde in phosphate buffer at pH 7 35. The use of osmium tetroxide as a fixative must, in general, be avoided for postembedding lectin histochemistry since it disturbs the reactivity of lectin binding sites.
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References
Herken, R, Fussek, M, and Thtes, M. (1988) Light and electron microscopical postembedding lectin histochemistry for WGA-binding sites in the renal cortex of the mouse embedded in polyhydroxy aromatic resins LR-White and LR-Gold Histochemutry 89, 277–282
Roth, J (1983) Application of lectin-gold complexes for electron microscopic localization of glycoconlugates on thin sections. J Histochem Cytochem 31, 987–999.
Robinson, D G, Ehlers, U, Herken, R., Herrmann, B, Mayer, F, and Schurmann, F-W (1987) Methods of Preparation for Electron Mcroscopy, Springer-Verlag, Berlin
Herken, R, Sander, B, and Hofmann, M. (1990) Ultrastructural localization of WGA, RCA I, LFA and SBA binding sites in the seven-day-old mouse embryo Histochemistry 94, 525–530
Salamat, M, Götz, W, Horster, A, Janotte, B, and Herken, R (1993) Ultrastructural locahzation of carbohydrates in Retchert’s membrane of the mouse Cell Tiss Res 272, 3775–381
Salamat, M., Gotz, W, Werner, J, and Herken, R (1993) Ultrastructural locahzation of lectin-binding sites in different basement membranes Histochem J 25, 464–468.
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© 1998 Humana Press Inc.
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Herken, R., Manshausen, B. (1998). Electron Microscopy. In: Rhodes, J.M., Milton, J.D. (eds) Lectin Methods and Protocols. Methods in Molecular Medicine™, vol 9. Humana Press. https://doi.org/10.1385/0-89603-396-1:111
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DOI: https://doi.org/10.1385/0-89603-396-1:111
Publisher Name: Humana Press
Print ISBN: 978-0-89603-396-2
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