脱钙处理对骨及其软组织免疫细胞化学染色的影响  

Effect of decalcification on immunohistochemical staining of osseous and soft tissues

作　　者：熊正文[1] 丁华野[2] 李春光[1] 田玉旺[2] 徐志勇[1] 

机构地区：[1]解放军251医院病理科,张家口075000 [2]北京军区总医院病理科

出　　处：《诊断病理学杂志》2000年第1期45-47,共3页Chinese Journal of Diagnostic Pathology

摘　　要：目的　探讨低温微波硝酸脱钙方法对骨及软组织抗原活性的影响。方法　根据实验条件分为 4组 :低温微波 10℃组 (Ⅰ组 ) ;冰箱 4℃ (Ⅱ组 ) ;室温 2 0℃ (Ⅲ组 ) ;及温箱 37℃ (Ⅳ组 )。分别使用 5 %硝酸、甲酸 -甲醛混合液、2 0 %甲酸液、Jenkins液、10 %乙烯二胺四乙酸钠 (EDTA)作为脱钙液。应用免疫组织化学LSAB染色和图像分析技术检测ANP ,5 HT ,S 10 0 ,NF ,GFAP和Vimentin的染色结果。结果　以硝酸作为脱钙剂时 ,Ⅰ组 (1.89± 0 97)阳性细胞的光密度值与Ⅱ组 (1 81± 1 0 2 )相近 ,均显著高于Ⅲ (1 0 3± 0 85 )、Ⅳ (0 76± 0 85 )组 ;脱钙液的温度越高 ,阳性细胞的光密度值越低 ,脱钙时间越短。低温微波硝酸与各种脱钙液的染色结果之间无明显差异 ,但脱钙时间显著缩短。结论　低温微波硝酸脱钙能够在较短时间内获得满意的染色效果。Objective To investigate the effect of decalcification by diffent agents with microwave (MW) at low temprature on the antigenicity of osseous and soft tissues. Methods According to the condition of decalcification, the experiments were divided into four groups: groupⅠwith MW at 10℃,groupⅡ,groupⅢ and groupⅣ at 4℃,20℃,and37℃,respectively without MW.Decalcification agents included 5% nitric acid (NA),fomic acid formalin(FAF),20% fomic acid (FA),Jenkins solution and 10% ethylene diamine tetracetic acid(EDTA).Expresstion of ANP,5 HT,S 100.NF,GFAP and vimentin was determined by immunohistochemical LSAB staining and image analysis. Results With NA as decalcifying agent, the optical dencity (OD)of positive cells in group`Ⅰ was similar to that in groupⅡand very signifycantly higher than those in groupⅢandⅣ.It showed the higher the temperature,the lower the OD of positive cells and the shorter the decalcifying time. No significant differences of OD existed among groups with different decalcifying agents described above ,but NA at 10℃with MW had shorted decalcifying time. Conclusion Very satisfactory staining results within relatively short time may be obtained with microwave treatment in nitric acid solation at low temperature for decalcification.

关 键 词：骨组织 脱钙 免疫组织化学 染色 

分 类 号：R681[医药卫生—骨科学]