快速脱钙骨组织病理切片台盼蓝-伊红染色方法研究

Trypan blue and eosin staining technique for rapid decalcification bone sections

目的研究建立台盼蓝-伊红(trypan blue and eosin,TE)染色法,并用于强酸脱钙后的骨组织病理切片染色,以提高制片质量。方法对10只大鼠胫骨进行5%硝酸脱钙、石蜡包埋、连续切片后,对每份胫骨标本分别进行苏木素-伊红(hematoxylin and eosin,HE)染色和甲苯胺蓝(toluidine blue,TB)染色及TE染色,比较不同染色方法的骨组织病理切片显微结构染色的差异。结果 10只大鼠胫骨组织标本每只分别进行3种染色,其中HE染色的骨小梁与骨髓分色不明显,骨细胞核无明显着色;TB染色的骨小梁与骨髓分色不明显,骨细胞边界模糊;TE染色的骨小梁显绛红色,骨髓细胞显紫红色,骨细胞陷窝、细胞核、骨小梁及附着的成骨细胞和破骨细胞、骨髓细胞中的巨噬细胞等结构特征清晰可辨。结论本研究所用的TE染色法适用于强酸脱钙处理的骨组织病理制片。

Objective To explore the feasibility of applying trypan blue and eosin staining （TE） technique to pathologic slices of bone tissue decalcified by 5% nitric acid. Method Tibias of 10 rats were decalcified by 5% nitric acid and then paraffin embedded sections were stained by hematoxylin and eosin （HE）, toluidine blue （TB） or TE. Results The color separation of bone trabecula and bone marrow was not obvious when using HE or TB. Addtionally, bone nucleus were not stained and the boundaries were hazy. In contrast, the trabecular and the bone marrow cells were stained deep-red and purplish red respecively when using TE method. The structure of osseous lacuna and bone cells include osteocyte, osteoblast, osteoclast and bone marrow macrophage is clear. Conclusion TE is suitable for pathologic slices of bone tissue decalcified by 5% nitric acid.