Summary
The detection of fibronectin (FN) in osmium-fixed and Araldite-embedded frog skin fragments was studied using a modification of Baskin's procedure (Baskin et al. 1979). Following the removal of Araldite from the semi-thin sections (0.5–1.0 μm) with ethanol-NaOH solution, the sections were bleached with hydrogen peroxide. FN was detected by indirect immunoperoxidase method. For precise localization of FN, careful attention was paid to the temperature, antibody concentrations and the quality of the ethanol-NaOH solution. Our results were in agreement with those that we had obtained previously for polyethylene glycol (PEG) sections, suggesting that the present procedure is useful for the detection of FN in Araldite-embedded biological specimens.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Andujar MB, Magloire H, Grimaud JA (1984) Fibronectin in basement membrane of Hertwig's epithelial sheat. Histochemistry 81:279–282
Asselineau D, Bernard BA, Bailly C, Darmon M, Prunieras M (1986) Human epidermis reconstructed by culture: Is it “normal”? J Invest Dermatol 86:181–186
Baskin DG, Erlandsen SL, Parsons JA (1979) Immunocytochemistry with osmium fixed tissue. I. Light microscopic localization of growth hormone and prolactin with the unlabeled antibodyenzyme method. J Histochem Cytochem 27:867–872
Clark RAF, Lanigan JM, Della Pelle P, Manseau E, Dvorak HF, Colvin RB (1982) Fibronectin and fibrin provide a provisional matrix for epidermal cell migration during wound reepithelialization. J Invest Dermatol 79:264–269
Denèfle JP (1973) Methode de culture organotypique de la peau de grenouille adulte (Rana esculenta). J Microscopie 17:97–102
Denèfle JP, Lechaire JP (1984) Epithelial locomotion and differentiation in frog skin cultures. Tissue Cell 16:499–517
Duband JL (1981) Matrice extracellulaire et morphogénèse. Thèse de 3ème cycle. Université Paris VII, pp. 18–19
Erlandsen SL, Parsons JA, Rodning CB (1979) Technical parameters of immuno-staining of osmicated tissue in epoxy sections. J Histochem Cytochem 27:1286–1289
Fujikawa LS, Forster CS, Gipson IK, Colvin RB (1984) Basement membrane components in healing rabbit corneal epithelial wounds: Immunofluorescence and ultrastructural studies. J Cell Biol 98:128–138
Grinnell F, Billingham RE, Burgess L (1981) Distribution of fibronectin during wound healing in vivo. J Invest Dermatol 76:181–189
Hogan DL, Smith GH (1982) Unconventional application of standard light and electron immunocytochemical analysis to aldehyde-fixed, araldite-embedded tissues. J Histochem Cytochem 30:1301–1306
Mauro A, Bertolotto A, Germano I, Giaccone G, Giordana MT, Migheli A, Schiffer D (1984) Collagenase in the immunohistochemical demonstration of laminin, fibronectin and factor VII/RAg in nervous tissue after fixation. Histochemistry 80:157–163
Repesh LA, Furcht LT, Smith D (1981) Immunocytochemical localization of fibronectin in limb tissue of the adult newt, Notophthalmus viridescens. J Histochem Cytochem 29:937–945
Zhu QL, Denèfle JP, Lechaire JP (1986) Fibronectin (FN) localizations in early wound healing of cultured frog skins. Biol Cell 57:161–168ff
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Zhu, Q.L. An immunocytochemical method for the localization of fibronectin in Araldite-embedded specimens. Histochemistry 86, 385–388 (1987). https://doi.org/10.1007/BF00494998
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00494998