LR-White and LR-Gold resins for postembedding immunofluorescence staining of laminin in mouse kidney
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This work describes a method for the immunolocalization of laminin on 1μm-thick tissue sections using a postembedding immunofluorescence technique. Embedding of unfixed or formaldehyde-fixed mouse renal cortex in either of the acrylic resins LR-White or LR-Gold permitted reliable postembedding immunofluorescence staining for laminin. LR-White was heat-cured at 50°C whereas LR-Gold was polymerized at −25°C. A stronger immunostaining for laminin was obtained from tissue embedded in polymerized LR-Gold compared with the staining from tissue embedded in LR-White. Prerequisites for adequate postembedding immunostaining are the partial dehydration of the tissue (maximum ethanol concentration, 70%) and pepsin treatment of the tissue sections prior to performing the immunostaining reactions.
KeywordsTissue Section Acrylic Resin Laminin Ethanol Concentration Immunofluorescence Staining
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- Barsky, S. H. Rao, N. C. Restrepo, C. &Liotta, L. A. (1984) Immunocytochemical enhancement of basement membrane antigens by pepsin: Applications in diagnostic pathology.Amer. J. Clin. Pathol. 82, 191–4.Google Scholar
- Craig, S. &Goodchild, D. J. (1982) Post-embedding immunolabelling. Some effects of tissue preparation on the antigenicity of plant proteins.Eur. J. Cell. Biol. 28, 251–6.Google Scholar
- Ekblom, P., Miettinen, M. Rapola, J. &Foidart, J-M. (1982) Demonstration of laminin, a basement membrane glycoprotein, in routinely processed formalinfixed human tissues.Histochemistry 75, 301–7.Google Scholar
- Herken, R., Fussek, M. & Thies, 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.Histochemistry (in press).Google Scholar
- Herken, R., Manshausen, B., Fussek, M. &Bonatz, G. (1987) Methodological dependence in the ultrastructural immunolocalization of laminin in tubular basement membranes of the mouse kidney.Histochemistry,87, 59–64.Google Scholar
- Newman, G. R., Jasani, B. &Williams, D. (1983) A simple post-embedding system for the rapid demonstration of tissue antigens under the electron microscope.Histochem. J. 15, 543–55.Google Scholar
- Timpl, R., Rohde, H., Robey, P. G., Rennard, S. J., Foidart, J-M. &Martin, G. R. (1979) Laminin — a glycoprotein from basement membranes.J. Biol. Chem. 254, 9933–7.Google Scholar
- Yokota, S. &Oda, T. (1985) Immunocytochemical demonstration of serine: Pyruvate aminotransferase in peroxisomes and mitochondria of rat kidney.Histochemistry 83, 81–5.Google Scholar