人类不脱钙骨组织的免疫组化与原位杂交

Immunohistochemistry and in situ hybridization technique for undecalcified bone sections of human

目的探讨在人类不脱钙骨组织切片上行免疫组化和原位杂交技术的可行性。方法采用Erben报道的改进的甲基丙稀酸甲酯低温塑料包埋方法包埋人类髂骨活检组织,制备不脱钙骨组织切片。切片行Goldners三色法及甲苯胺蓝染色观察骨组织的形态结构;采用免疫组化SP法和原位杂交技术检测骨组织中碱性成纤维细胞生长因子(FGF2)蛋白和mRNA的表达。结果组织形态学上,两种染色法均示骨组织中成骨细胞、破骨细胞及骨细胞清晰可辨,钙化的骨基质与类骨质分界清楚。免疫组化和原位杂交示FGF2蛋白和mRNA表达于骨髓腔中一些单个核细胞及少许成骨细胞的胞浆中。此外,骨髓基质亦可见FGF2蛋白和mRNA的阳性表达。结论低温塑料包埋方法制备的人类不脱钙骨组织切片可同时进行免疫组化和原位杂交的检测观察。

Objective To explore the feasibility of applying immunohistochemistry （IH）and in situ hybridization （ISH） technique to the undecalcified bone sections of human. Methods Two iliac creast biopsies were embedded with the methylmethacrylate（MMA）low-temperature plastic embedding method reported by Erben and then made into undecalcified sections. These sections were observed in morphology by Goldner＇s Masson Trichrome staining and toluidine blue staining. FGF-2 protein and mRNA in the bone sections were detected by IH and ISH, Results Both Goldner＇s Masson Trichrome staining and toluidine blue staining showed the cells in bone tissue such as osteoblast, osteoclast and osteocyte were clearly seen, and osteoid were easily distinguished from mature bone tissue. The positive signals of FGF-2 protein and mRNA were found in the cytoplasm of some mononuclear ceils in the bone marrow and osteoblast, the matrix in the bone marrow was positively stained, too. Conclusions The undecalcified bone sections of human made in the method can satisfy the need of IH and ISH.