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中华临床实验室管理电子杂志

中华临床实验室管理电子杂志 ›› 2017, Vol. 05 ›› Issue (03) : 171 -175. doi: 10.3877/cma.j.issn.2095-5820.2017.03.010

所属专题： 文献；

实验研究

快速脱钙骨组织病理切片台盼蓝-伊红染色方法研究
刘洋1, 柯诗韵2, 彭方美2, 翟玉莹2, 肖珊2, 叶小天2, 陈珺2, 陆幸妍2,†()   
  1. 1. 510663　广州，广东省食品药品职业技术学校
    2. 510006　广州，广东药科大学生命科学与生物制药学院/广东省生物技术候选药物研究重点实验室
  • 收稿日期:2017-04-30 出版日期:2017-08-28
  • 通信作者: 陆幸妍
  • 基金资助:
    广东省科技计划项目资助课题(2013B060300022，2014A020212241)

Trypan blue and eosin staining technique for rapid decalcification bone sections

Yang Liu1, Shiyun Ke2, Fangmei Peng2, Yuying Zhai2, Shan Xiao2, Xiaotian Ye2, Jun Chen2, Xingyan Lu2,†()   

  1. 1. Guangdong Food and Drug Vocation-technical School, Guangzhou 510663, China
    2. School of Life Science and Biopharmacy, Guangdong Pharmaceutical University/ Guangdong Provincial Key Laboratory of Biotechnology Candidate Drug Research, Guangzhou 510006, China
  • Received:2017-04-30 Published:2017-08-28
  • Corresponding author: Xingyan Lu
  • About author:
    Corresponding author: Lu Xingyan, Email:
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引用本文：

刘洋, 柯诗韵, 彭方美, 翟玉莹, 肖珊, 叶小天, 陈珺, 陆幸妍. 快速脱钙骨组织病理切片台盼蓝-伊红染色方法研究[J/OL]. 中华临床实验室管理电子杂志, 2017, 05(03): 171-175.

Yang Liu, Shiyun Ke, Fangmei Peng, Yuying Zhai, Shan Xiao, Xiaotian Ye, Jun Chen, Xingyan Lu. Trypan blue and eosin staining technique for rapid decalcification bone sections[J/OL]. Chinese Journal of Clinical Laboratory Management(Electronic Edition), 2017, 05(03): 171-175.

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。

关键词: 快速脱钙, 苏木素-伊红染色, 甲苯胺蓝染色, 台盼蓝-伊红染色
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.

Key words: Rapid decalcification, Hematoxylin-eosin staining, Toluidine blue staining, Trypan blue and eosin staining
图1 大鼠胫骨上段切片HE染色图(4μm, ×20)
图2 大鼠胫骨上段切片TB染色图(4μm, ×20)
图3 大鼠胫骨上段切片TE染色图(4μm, ×20)
图4 大鼠骨组织细胞HE染色图(4μm, ×200)
图5 大鼠骨组织细胞TB染色图(4μm, ×200)
图6 大鼠骨组织细胞TE染色图(4μm, ×200)
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