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Journal of Histochemistry and Cytochemistry, Vol. 51, 61-67, January 2003, Copyright © 2003, The Histochemical Society, Inc.

ARTICLE

Quantitative Immunogold Labeling of Bone Sialoprotein and Osteopontin in Methylmethacrylate-embedded Rat Bone

O. Labouxa,d, L-G. Ste-Marieb, F.H. Glorieuxc, and A. Nancia
a Laboratory for the Study of Calcified Tissues and Biomaterials, Faculty of Dentistry, Université de Montréal, Québec, Canada
b Metabolic Bone Diseases Laboratory, Centre de Recherche du CHUM, Hôpital St Luc, Montréal, Québec, Canada
c Genetics Unit, Shriners Hospital for Children, and Departement of Surgery and Human Genetics, McGill University, Montreal, Québec, Canada
d INSERM E.M.I., Nantes, France

Correspondence to: A. Nanci, Université de Montréal, Faculty of Dentistry, Dept. of Stomatology, PO Box 6128, Station Centre-Ville, Montreal, QC, Canada H3C 3J7. E-mail: Antonio.Nanci@UMontreal.CA

Methylmethacrylate (MMA) embedding of undecalcified bone is routinely employed for histomorphometric analyses. Although MMA-embedded bone has been used for immunolabeling at the light microscopic level after removal of the resin, there are no such reports for electron microscopy. The aim of the present study was to determine whether MMA embedding can be used for ultrastructural immunolabeling and how it compares to LR White (LRW), an acrylic resin frequently used for immunocytochemistry of bone. Rat tibiae were fixed by vascular perfusion with aldehyde and embedded either in MMA or LRW resin. Thin sections were processed for postembedding protein A–gold immunolabeling with antibodies to rat bone sialoprotein (BSP) and osteopontin (OPN). The density of gold particles over bone was quantified. The density and distribution of immunolabeling for BSP and OPN respectively, were comparable between MMA and LRW. These results indicate that MMA performs as well as LRW for the ultrastructural immunolabeling of noncollagenous bone matrix proteins. (J Histochem Cytochem 51:61–67, 2003)

Key Words: bone, bone sialoprotein, immunocytochemistry, LR White resin, methylmethacrylate osteopontin


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