免疫染色的應(yīng)用了解細(xì)胞來源課件

口

腔

病

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科免疫組織化學(xué)染色介紹AnIntroductionofImmunocytochemistry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(yīng)用口腔病理科免疫組織化學(xué)染色介紹AnIntroducImmunohistochemistrycytochemistryAntibodyAntigenAntibodyAntigenDetectionSystemAntigenAntibody釣魚作為比喻Antibody魚餌魚Antigen浮標(biāo)LabellingSystem釣竿MethodMethodFixation/FreshLabellingSystemPolyclonal-amixofIg(rabbit)Monoclonal-asingleclassofIg(mouse)Synthetic\CommercialEnzyme:HorseradishPeroxidase(HP),AlkalinePhosphatase(AP);FITC,TRITC(tetramethylrhadamineisothiocyanate);gold;isotope

Chromagen:Diaminobenzidine(DAB)(Autolysis)24hExcessiveEnzyme(trypsin)MicrowaveAutoclave免疫染色的原理ImmunohistochemistryAntibodyAnMeritsSimpleQuickCheapDrawbackHighbackgroundLowerintensityDirectMethodAntigenCytoplasmAntigenEndothelialcellNucleusLabelling(Onlyone)LabelledPrimaryAntibodyHumanRabbit/mouseanti-human2.

免疫染色的種類MeritsDrawbackLowerintensityDHigherintensityMeritsSimpleQuickCheapDrawbackHighbackgroundIndirectMethodCytoplasmAntigenAntigenEndothelialcellNucleusUnlabelledPrimaryAntibodyFirstlabellingSecondlabellingHumanRabbitanti-humanHorseanti-rabbitLabelledSecondaryAntibodyHigherintensityMeritsDrawbackPAPMethodAntigenCytoplasmAntigenEndothelialcellNucleusUnlabelledSecondary(Bridging)AntibodyPeroxidaseAnti-Peroxidase(PAP)UnlabelledPrimaryAntibodyFirstlabellingSecondlabellingThirdlabellingHigherintensityDrawback

ExpensiveTimeconsumingHumanRabbitanti-humanHorseanti-rabbitRabbitPAPPAPMethodAntigenCytoplasmAntABCMethodAntigenCytoplasmAntigenEndothelialcellNucleusUnlabelledPrimaryAntibodyHigherintensityDrawback

ExpensiveHumanRabbitanti-humanHorseanti-rabbitBiotinylatedSecondary(Bridging)AntibodyAvidin-BiotinylatedPeroxidaseComplex(ABC)ABCMethodAntigenCytoplasmAntCytoplasmNucleusAntigenAntigenEndothelialcellEliminationofEndogenousPeroxidaseLabelledPrimaryAntibodyEndogenousperoxidaseH2O2H2O2H2O2H2O2EliminateendogeousperoxidaseLabellingsystem:PeroxidaseDrawback

HighbackgroundCytoplasmNucleusAntigenAntigenCytoplasmNucleusEliminationofNon-SpecificStainingAntigenAntigenEndothelialcellAntigenAntigenEndothelialcellLabelledPrimaryAntibodyLabelledPrimaryAntibodyNormalSerum(samespeciesof2ndAb)EndothelialcellAntigenCollagenEndothelialcellLabelledPrimaryAntibodyLabelledPrimaryAntibodySpecificbindingNon-specificbindingCytoplasmNucleusEliminationofAvidin-Biotin

Peroxidase

Complex

(ABC)

methodH2O2Normalserum1oAbBiotinylated2oAbABCreagentDABstainHsuSM,1981Avidin-BiotinPeroxidaseComplPAPMethodAntigenAntigenLymphoidcellNucleusUnlabelledPrimaryAntibodyHumanRabbitanti-humanHorseanti-rabbitRabbitAPAAPUnlabelledSecondary(Bridging)Antibody(1)DoubleStainingCytoplasmUnlabelledSecondary(BridgingAntibody(2)Rabbit

PAPAPAAPPAPAlkalinePhosphasteAntiAlkalinePhosphastePAPMethodAntigenAntigenLymphComparisonofdifferentmethodsofIHCMethodsMeritsDrawbacksDirectmethodSimpleQuick

CheapHighbackgroundIndirectmethodSimpleQuickCheapHighbackgroundPAPHighintensityExpensiveTimeconsumingABCHighintensityExpensiveComparisonofdifferentmethod3.

免疫染色的應(yīng)用了解細(xì)胞來源3.免疫染色的應(yīng)用了解細(xì)胞來源3.

免疫染色的應(yīng)用了解細(xì)胞來源3.免疫染色的應(yīng)用了解細(xì)胞來源3.

免疫染色的應(yīng)用了解細(xì)胞來源3.免疫染色的應(yīng)用了解細(xì)胞來源3.

免疫染色的應(yīng)用了解細(xì)胞來源3.免疫染色的應(yīng)用了解細(xì)胞來源3.

免疫染色的應(yīng)用了解細(xì)胞來源3.免疫染色的應(yīng)用了解細(xì)胞來源3.

免疫染色的應(yīng)用了解細(xì)胞來源3.免疫染色的應(yīng)用了解細(xì)胞來源3.

免疫染色的應(yīng)用了解細(xì)胞來源A:CK(AE1/AE3)showsdiffusecytoplasmicstainingpatternatfullthicknessofthesquamousmucosa.

B:p63showsnuclearstainingpatternatbasaltointermediatelayerofsquamousmucosa.C:EMAshowsmembranousstainingpatternofepithelialcells.D:Vimentinshowsdiffusecytoplasmicstainingpatternonsubepithelialstromalcells.TheepitheliumisnegativeforvimentinOralmucosa(100X)3.免疫染色的應(yīng)用了解細(xì)胞來源A:CK(AE1/AE33.

免疫染色的應(yīng)用了解細(xì)胞來源C(SmoothMuscleActin,SMA)stainontumorcellswithmyoepithelialdifferentiation.Pleomorphicadenomaofparotidgland(400X)A:CK(AE1/AE3)stainsontumorcellswithductaldifferentiation.B(S-100protein)ontumorcellswithmyoepithelialdifferentiation.3.免疫染色的應(yīng)用了解細(xì)胞來源C(SmoothMusc3.

免疫染色的應(yīng)用了解細(xì)胞來源D:Vimentinshowscytoplasmicstainingpatternonsarcomatoidcarcinoma.Neuroendocrinecarcinomaofbreast(400X).E(chromogranin)andF(synaptophysin)showgranularcytoplasmicstainingpatternontumorcells.3.免疫染色的應(yīng)用了解細(xì)胞來源D:Vimentinsh3.

免疫染色的應(yīng)用了解細(xì)胞來源Metastaticovariancolonictypemucinouscarcinoma(400X).Carcinoembronicantigen(CEA)stainsonapicalsurfaceofadenocarcinoma.Esophagealmoderatelydifferentiatedsquamouscellcarcinoma(400X).p16showsstrongfullthicknessoverexpressiononthecytoplasmandnucleusoftumorcells.Theadjacentmucousglandisnegativeforp16.3.免疫染色的應(yīng)用了解細(xì)胞來源Metastaticova3.

免疫染色的應(yīng)用了解細(xì)胞來源Ovarianintestinaltypemucinousadenocarcinoma(400X).I(CK7)andJ(CK20)coexpressontumorwithheterogenouscytoplasmicstainingpattern.3.免疫染色的應(yīng)用了解細(xì)胞來源Ovarianintest3.

免疫染色的應(yīng)用了解細(xì)胞來源K:Colonicadenocarcinoma(400X).CK20showsdiffuseandstrongcytoplasmicstainingpatternoftumorcells.L:Malignantmelanoma(400X).HMB-45showscytoplasmicstainingpatternonmelanomacells.3.免疫染色的應(yīng)用了解細(xì)胞來源K:Colonicade3.

免疫染色的應(yīng)用了解細(xì)胞來源N(CD15)andO(CD30)coexpressoncellmembraneand