微波-EDTA脱钙技术在免疫组织化学染色中的应用研究

Application of decalcification technique with microwave and ethylene diamine tetracetic acid to immunohistochemistry

目的 探讨微波 (MW ) 乙烯二胺四乙酸钠 (EDTA)快速脱钙技术对骨及软组织抗原活性的影响。 方法 根据实验条件将待脱钙组织分为 4组 ,即MW EDTA(15℃ ) ,Jenkins脱钙 固定液 (2 0℃ ) ,10 %EDTA(2 0℃ )和 10 %硝酸 (2 0℃ ) ,其中MW EDTA方法中EDTA的浓度分别是 4%、7%、10 %、13 %和 16%。应用免疫组织化学LSAB染色 ,光镜观察免疫组织化学染色结果。 结果 MW EDTA脱钙的免疫组织化学染色阳性细胞显著高于Jenkins脱钙 固定液 (2 0℃ )、10 %EDTA(2 0℃ )及 10 %硝酸脱钙 (2 0℃ ) 3种脱钙方法 ,其时间明显缩短。其中用 10 %和 13 %浓度的EDTA脱钙的组织 ,其免疫组织化学染色阳性细胞又明显多于EDTA浓度为 4%、7%和 16%者。 结论 以MW 10 %EDTA技术进行组织脱钙 ,其脱钙时间短。

Objective To investigate the effect of rapid decalcification techniques with microwave(MW) at low temperature and ethylene diamine tetracetic acid(EDTA) on the antigenicity of osseous and soft tissues. Methods All tissues were distributed into four groups and each group was treated with one of the following four decalcification techniques,MW EDTA(15℃), Jenkin's decalcificative fixative solution,10% EDTA(20℃) and 10% nitric acid(20℃). In the MW EDTA group, the concentration of EDTA had 5 options,4%, 7%, 10%, 13% and 16%. Immunohistochemistry method LSAB was performed on sections of the tissues.The staining was observed under light microscope. Results The number of positive cells was much greater in MW DETA group than those in the groups processed with the other solutions,MW EDTA also took shorter time.Of the differnt concentrations of EDTA, 10% and 13% resulted in better staining than the other three concentrations. Conclusions MW 10% EDTA solution could obtain satisfying immunohistochemical staining with relatively shorter time.