病理学技术原的应用.pptVIP

试验片：+ 阳性对照片：+ 阴性对照片：+ 空白对照片：- 结果：可疑阳性 原因： 阴性对照组织选择不准确，含有靶抗原 试验片：+ 阳性对照片：+ 阴性对照片：- 空白对照片：- 结果：阳性 原因： 方法可靠，实验片含靶抗原。 阴性对照切片-，阳性对照标本和待检测切片呈弱阳性 原因： 标本固定不当 抗体试剂问题：浓度、效价低 抗原修复不当 显色时间短 切片染色深或整张切片出现染色 抗体浓度过高 切片显色时间过长 组织没有正确封闭 冲洗不够充分 切片出现非特异性背景染色 内源性过氧化物酶、内源性生物素过多 血清封闭时间过短 抗体不纯 组织片内出血或坏死成分 冲洗不彻底 四、免疫荧光技术 将免疫学方法(抗原抗体特异结合)与荧光标记技术结合起来研究特异蛋白抗原在细胞内分布的方法。由于荧光素所发的荧光可在荧光显微镜下检出，从而可对 抗原进行细胞定位 免疫荧光技术 五、分子生物学技术—原位杂交 分子生物学技术 —原位杂交和原位PCR HPV DNA CK 组织芯片 HE 组织芯片 免疫学 特染 病理学技术 杂交 Autopsy: verity of disease, reason of, death,response to treatment.the differences among surgeon,physician and pathologist! Biopsy: verity of disease, response to treatment, sometimes hard to be receptible by patient frozen sections:quickly meet the needs during the operation,special histochemistry Cytology:simple and convenient to perform,easy acceptable with low expense.tissue culture,pathogen identifying restrictions in final diagnosis Animal model:pathogenesis,influence of treatment Tissue cell culture:pure cells from one ancestor,easy to analysis result. Methodology Qualitative and quantitative methods,select depend on different object In Situ Hybridization techniques allow the demonstration of specific nucleic acid sequences (genes) in their cellular environment. Above: Human Papillomavirus DNA demonstrated by In Situ Hybridization (pink)in epithelial cells identified by indirect immunofluorescence using antibody against cytokeratin (green Human Papillomavirus DNA Double staining for EBER and a second antibody Double staining after in situ hybridization for EBER can be done, but the incubation in denaturing conditions with the probe masks numerous antigens, in an irreversible way. You should pre-test your second primary antibody on a section treated as if you would have done ISH. You can use an incubation with 50% Dimethylformamide for 30 min. to test your reagents. Double staining can then be done as detailed 组织芯片技术可以将数十个甚至上千个不同个体的临床组织标本按预先设计的顺序排列在一张玻片进行分析研究，是一种高通量、多样本的分析工具。它使科研人员第一次