Evaluation of Decalcification Techniques for Rat Femurs Using HE and Immunohistochemical Staining.
Evaluation of Decalcification Techniques for Rat Femurs Using HE and Immunohistochemical Staining.
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HE 和免疫组织化学染色评价大鼠股骨脱钙技术
DOI：
10.1155/2017/9050754
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发表时间：
2017
影响因子：
--
通讯作者：
Gao S
中科院分区：
文献类型：
--
作者：
Liu H;Zhu R;Liu C;Ma R;Wang L;Chen B;Li L;Niu J;Zhao D;Mo F;Fu M;Brömme D;Zhang D;Gao S
瞄准在常规组织病理学中，脱钙是矿化组织的重要步骤。本研究旨在评价不同脱钙液对SD大鼠股骨形态学和抗原性保存的影响。材料和方法。采用10%中性缓冲EDTA、3%硝酸、5%硝酸和8%盐酸/甲酸四种不同的脱钙溶液去除大鼠股骨中的矿物质。采用振荡和低温处理样品。HE染色和免疫组化染色观察骨形态和抗原性。主要发现。在病理检查中，不同的脱钙液可能会影响石蜡包埋切片的形态学和染色质量。四种脱钙液中，3%硝酸脱钙效果最好。10%中性缓冲EDTA和5%硝酸是IHC染色的首选脱钙剂。意义本研究探讨了不同脱钙剂对骨结构和抗原性的保护作用，这将有助于制定合适的协议，骨组织的分析。
Aim. In routine histopathology, decalcification is an essential step for mineralized tissues. The purpose of this study is to evaluate the effects of different decalcification solutions on the morphological and antigenicity preservation in Sprague Dawley (SD) rat femurs. Materials and Methods. Four different decalcification solutions were employed to remove the mineral substances from rat femurs, including 10% neutral buffered EDTA, 3% nitric acid, 5% nitric acid, and 8% hydrochloric acid/formic acid. Shaking and low temperature were used to process the samples. The stainings of hematoxylin-eosin (HE) and immunohistochemical (IHC) were employed to evaluate the bone morphology and antigenicity. Key Findings. Different decalcification solutions may affect the quality of morphology and the staining of paraffin-embedded sections in pathological examinations. Among four decalcifying solutions, 3% nitric acid is the best decalcifying agent for HE staining. 10% neutral buffered EDTA and 5% nitric acid are the preferred decalcifying agents for IHC staining. Significance. The current study investigated the effects of different decalcifying agents on the preservation of the bone structure and antigenicity, which will help to develop suitable protocols for the analyses of the bony tissue.