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Bone samples were embedded undecalcified.
Bone samples were embedded in paraffin and 5 µm sections were stained for tartrate-resistant acid phosphatase (TRAP).
Bone samples were embedded in acrylic resin based on methyl methacrylate as described before [ 4, 5, 16, 17].
While the implants were removed, the peri-implant bone samples were embedded with paraffin wax and cut into 3 μm thick sections, along the longitudinal implant axis, using a motorized microtome.
Using the method of Donath and Breuner 1982 [ 12] undecalcified bone samples were embedded in Technovit 9100® New (Heraeus Kulzer, Hanau, Germany) – a low temperature methylmethacrylate embedding system that significantly improves tissue antigenicity preservation allowing polymerisation at -20°C and quantitative assessment of bone-matrix proteins.
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Then, this bone sample was embedded in paraffin.
Before cutting the final slices of bone the samples were embedded in acrylic resin [ 21].
Spleen and bone marrow samples were embedded in O.C.T. compound medium (Sakura Alphen aan den Rijn, The Netherlands) in cryomold vessels, snap frozen in liquid nitrogen for 10 min and stored at −80 °C.
After separation of the calvarial bones, the samples were embedded in paraffin blocks.
After separation of the parietal bones through the midline sagittal suture, the calvarial samples were embedded in paraffin blocks.
Bone samples were decalcified for 3 weeks in 10 % EDTA (pH 7.4) with weekly changes, and then all the samples were embedded in paraffin.
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