脱钙处理对骨及其软组织免疫细胞化学染色的影响
作者：熊正文 丁华野 李春光 田玉旺 徐志勇
单位：熊正文(解放军251医院病理科 张家口 075000)；丁华野(北京军区总医院病理科)；李春光(解放军251医院病理科 张家口 075000)；田玉旺(北京军区总医院病理科)；徐志勇(解放军251医院病理科 张家口 075000)
关键词：骨组织；脱钙方法；硝酸；微波；免疫组织化学
诊断病理学杂志000116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20℃(Ⅲ组)；及温箱37℃(Ⅳ组)。分别使用5%硝酸、甲酸-甲醛混合液、20%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-100，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±1.02)相近，均显著高于Ⅲ(1.03±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。
, 百拇医药
分类号：R36 文献标识码：A
文章编号：1007-8096(2000)01-0045-03
Effect of decalcification on immunohistochemical staining of osseous and soft tissues
Xiong Zhengwen,Ding huaye,Li Chunguang
(Department of Pathology, 251 Hospital of PLA, Zhangjiakou 075000)
Abstract：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 divid-ed into four groups: group Ⅰ with MW at 10℃ ,group Ⅱ ,groupⅢ and group Ⅳ 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 im-munohistochemical LSAB staining and image analysis. Results With NA as decalcifying agent, the optical dencity (OD)of pos-itive cells in group‘Ⅰ was similar to that in group Ⅱ and very signifycantly higher than those in groupⅢ andⅣ. It showed the high-er 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. Con-clusion Very satisfactory staining results within relatively short time may be obtained with microwave treatment in nitric acid sola-tion at low temperature for decalcification.
, http://www.100md.com
Keywords：Osseous tissue Decalcification Nitric acid Microwave Immunnohistochemistry
参考文献：
[1]蔡文琴，王伯云主编.实用免疫细胞在化学与核酸分子杂交技术.成都：四川科学技术出版社，1994.1-20
[2]Piffko J，Bankfalvi A，Ofiner D，et al.Immunohistochemical detection of p53 protein in archival tissues from squamous cell carcinomas of the oral cavity using wet autoclave antigen retrieval.J phthol,1995，176：69-75
, 百拇医药 [3]YM YJ，Anniko M.Distribution of alpha-ANP in the cochlea and the vestibular organs.ORL，1994,56：73-7
[4]陈合新，邱建华，乔莉，等.豚鼠血管纹心钠素(ANP)免疫组化分布超微定位.中华耳鼻咽喉科杂志，1993，28：339
[5]Yoon YJ，Hellstrom S.Immnuohistochemical localizatio of alpha-atrial na-triuretie polypeptide in the rat cochlea.Acta otolaryngol(Stockh)，1992，112：604-10
[6]Shi SR，Imam A，Young L，et al.Antigen retrieval immunohistochemistry under the in fluence of pH using monoclonal antibodies.J Histochem Cy-tochem，1995，43：193-201
[7]田玉旺，丁华野，邓永江，等.免疫组织化学反应产物图像定量分析技术.细胞与分子免疫学杂志，1997，13(S1)：63
收稿日期：1998-09-07
修稿日期：1999-05-04, 百拇医药
单位：熊正文(解放军251医院病理科 张家口 075000)；丁华野(北京军区总医院病理科)；李春光(解放军251医院病理科 张家口 075000)；田玉旺(北京军区总医院病理科)；徐志勇(解放军251医院病理科 张家口 075000)
关键词：骨组织；脱钙方法；硝酸；微波；免疫组织化学
诊断病理学杂志000116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20℃(Ⅲ组)；及温箱37℃(Ⅳ组)。分别使用5%硝酸、甲酸-甲醛混合液、20%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-100，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±1.02)相近，均显著高于Ⅲ(1.03±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。
, 百拇医药
分类号：R36 文献标识码：A
文章编号：1007-8096(2000)01-0045-03
Effect of decalcification on immunohistochemical staining of osseous and soft tissues
Xiong Zhengwen,Ding huaye,Li Chunguang
(Department of Pathology, 251 Hospital of PLA, Zhangjiakou 075000)
Abstract：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 divid-ed into four groups: group Ⅰ with MW at 10℃ ,group Ⅱ ,groupⅢ and group Ⅳ 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 im-munohistochemical LSAB staining and image analysis. Results With NA as decalcifying agent, the optical dencity (OD)of pos-itive cells in group‘Ⅰ was similar to that in group Ⅱ and very signifycantly higher than those in groupⅢ andⅣ. It showed the high-er 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. Con-clusion Very satisfactory staining results within relatively short time may be obtained with microwave treatment in nitric acid sola-tion at low temperature for decalcification.
, http://www.100md.com
Keywords：Osseous tissue Decalcification Nitric acid Microwave Immunnohistochemistry
参考文献：
[1]蔡文琴，王伯云主编.实用免疫细胞在化学与核酸分子杂交技术.成都：四川科学技术出版社，1994.1-20
[2]Piffko J，Bankfalvi A，Ofiner D，et al.Immunohistochemical detection of p53 protein in archival tissues from squamous cell carcinomas of the oral cavity using wet autoclave antigen retrieval.J phthol,1995，176：69-75
, 百拇医药 [3]YM YJ，Anniko M.Distribution of alpha-ANP in the cochlea and the vestibular organs.ORL，1994,56：73-7
[4]陈合新，邱建华，乔莉，等.豚鼠血管纹心钠素(ANP)免疫组化分布超微定位.中华耳鼻咽喉科杂志，1993，28：339
[5]Yoon YJ，Hellstrom S.Immnuohistochemical localizatio of alpha-atrial na-triuretie polypeptide in the rat cochlea.Acta otolaryngol(Stockh)，1992，112：604-10
[6]Shi SR，Imam A，Young L，et al.Antigen retrieval immunohistochemistry under the in fluence of pH using monoclonal antibodies.J Histochem Cy-tochem，1995，43：193-201
[7]田玉旺，丁华野，邓永江，等.免疫组织化学反应产物图像定量分析技术.细胞与分子免疫学杂志，1997，13(S1)：63
收稿日期：1998-09-07
修稿日期：1999-05-04, 百拇医药