Bain S D, Impeduglia T M, Rubin C T
Department of Orthopaedics, State University of New York, Stony Brook 11794-8181.
Stain Technol. 1990;65(4):159-63. doi: 10.3109/10520299009108065.
A technique for demonstrating cement lines in thin, undecalcified transverse sections of cortical bone has been developed. Cortical bone samples are processed and embedded undecalcified in methyl methacrylate plastic. After sectioning at 3-5 microns, cross-sections are transferred to a glass slide and flattened for 10 min. Sections of cortical bone are stained for 20 sec free-floating in a fresh solution of 1% toluidine blue dissolved in 0.1% formic acid. The section is dehydrated in t-butyl alcohol, cleared in xylene, and mounted with Eukitt's medium. Reversal lines appear as thin, scalloped, dark blue lines against a light blue matrix, whereas bone formation arrest lines are thicker with a smooth contour. With this technique cellular detail, osteoid differentiation, and fluorochrome labels are retained. Results demonstrate the applicability of a one-step staining method for cement lines which will facilitate the assessment of bone remodeling activity in thin sections of undecalcified cortical bone.
已开发出一种在皮质骨薄的、未脱钙横切面上显示黏固线的技术。将皮质骨样本进行处理并以未脱钙状态包埋于甲基丙烯酸甲酯塑料中。在切成3 - 5微米厚的切片后，将横截面转移至载玻片上并展平10分钟。将皮质骨切片在新鲜配制的、溶解于0.1%甲酸中的1%甲苯胺蓝溶液中自由漂浮染色20秒。切片在叔丁醇中脱水，在二甲苯中透明，并用尤基特介质封固。反转线呈现为浅蓝色基质背景上的细的、扇形的深蓝色线条，而骨形成停滞线更厚且轮廓光滑。通过该技术，细胞细节、类骨质分化和荧光染料标记得以保留。结果表明一种用于黏固线的一步染色方法的适用性，这将有助于评估未脱钙皮质骨薄片中的骨重塑活性。
