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1、口 腔 病 理 科,免疫組織化學染色介紹,An IntroductionofImmunocytochemistry,歡迎出席本課程,明白免疫染色的原理 瞭解免疫染色的種類 知道免疫染色的應用,DetectionSystem,Antigen,Antibody,釣魚作為比喻,Method,Fixation/Fresh,Labelling System,Enzyme:Horseradish Peroxidase(HP),Alkaline Phosphatase(AP);FITC,TRITC(tetramethyl rhadamineisothiocyanate);gold;isotope,Chrom
2、agen:Diamino benzidine(DAB),Enzyme(trypsin)MicrowaveAutoclave,免疫染色的原理,Merits Simple Quick Cheap,Drawback High background,Lower intensity,Labelling(Only one),Human,Rabbit/mouseanti-human,2.免疫染色的種類,Higher intensity,Merits Simple Quick Cheap,Drawback High background,Indirect Method,Cytoplasm,First labe
3、lling,Second labelling,Human,Rabbitanti-human,Horseanti-rabbit,Labelled SecondaryAntibody,PAP Method,Antigen,Cytoplasm,Antigen,Endothelial cell,UnlabelledSecondary(Bridging)Antibody,PeroxidaseAnti-Peroxidase(PAP),First labelling,Second labelling,Third labelling,Higher intensity,Drawback Expensive Ti
4、me consuming,Human,Rabbitanti-human,Horseanti-rabbit,RabbitPAP,ABC Method,Antigen,Cytoplasm,Antigen,Endothelial cell,Higher intensity,Drawback Expensive,Human,Rabbitanti-human,Horseanti-rabbit,BiotinylatedSecondary(Bridging)Antibody,Cytoplasm,Elimination of Endogenous Peroxidase,Endogenousperoxidase
5、,Labelling system:Peroxidase,Drawback High background,Cytoplasm,Elimination of Non-Specific Staining,Specific binding,Non-specific binding,Avidin-Biotin Peroxidase Complex(ABC)method,H2O2,Normal serum,1o Ab,Biotinylated 2o Ab,ABCreagent,DABstain,Hsu SM,1981,PAP Method,Antigen,Antigen,Lymphoid cell,H
6、uman,Rabbitanti-human,Horseanti-rabbit,Rabbit APAAP,UnlabelledSecondary(Bridging)Antibody(1),Double Staining,Cytoplasm,UnlabelledSecondary(BridgingAntibody(2),Rabbit PAP,APAAP,PAP,Alkaline Phosphaste AntiAlkaline Phosphaste,Comparison of different methods of IHC,3.免疫染色的應用,了解細胞來源,3.免疫染色的應用,了解細胞來源,3.免
7、疫染色的應用,了解細胞來源,3.免疫染色的應用,了解細胞來源,3.免疫染色的應用,了解細胞來源,3.免疫染色的應用,了解細胞來源,3.免疫染色的應用,了解細胞來源,A:CK(AE1/AE3)shows diffuse cytoplasmic staining pattern at full thickness of the squamous mucosa.B:p63 shows nuclear staining pattern at basalto intermediate layer of squamous mucosa.C:EMA shows membranous staining pat
8、tern of epithelial cells.D:Vimentin shows diffuse cytoplasmic staining pattern on subepithelialstromal cells.The epithelium isnegative for vimentin,Oral mucosa(100X),3.免疫染色的應用,了解細胞來源,C(Smooth Muscle Actin,SMA)stain on tumor cells with myoepithelialdifferentiation.,Pleomorphic adenoma of parotid glan
9、d(400X),A:CK(AE1/AE3)stains on tumor cells with ductal differentiation.,B(S-100 protein)on tumor cells with myoepithelialdifferentiation.,3.免疫染色的應用,了解細胞來源,D:Vimentin shows cytoplasmic staining pattern on sarcomatoid carcinoma.,Neuroendocrine carcinoma of breast(400X).E(chromogranin)andF(synaptophysi
10、n)show granularcytoplasmic staining pattern on tumor cells.,3.免疫染色的應用,了解細胞來源,Metastatic ovarian colonic type mucinous carcinoma(400X).Carcinoembronic antigen(CEA)stains on apical surface of adenocarcinoma.,Esophageal moderately differentiatedsquamous cell carcinoma(400X).p16 shows strong full thickn
11、ess overexpression on the cytoplasm andnucleus of tumor cells.The adjacent mucous gland is negative for p16.,3.免疫染色的應用,了解細胞來源,Ovarian intestinal type mucinous adenocarcinoma(400X).I(CK7)and J(CK20)coexpress on tumor with heterogenouscytoplasmic staining pattern.,3.免疫染色的應用,了解細胞來源,K:Colonic adenocarci
12、noma(400X).CK20 shows diffuse and strongcytoplasmic staining pattern of tumor cells.,L:Malignant melanoma(400X).HMB-45 shows cytoplasmic staining pattern on melanoma cells.,3.免疫染色的應用,了解細胞來源,N(CD15)and O(CD30)coexpress on cell membrane and paranuclearspace of RS cells,Nodular sclerosing type of classical Hodgkins lymphoma,M:LCA stains on all lymphoid cells including large malignant RS cells and surround reactive small lymphocytes.,3.免疫染色的應用,了解細胞來源,-,
