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1、眼科研究2010年1月第28卷第1期15实验研究不同抗原修复法对视网膜石蜡切片相关功能蛋白表达检测的影响李颖胡运韬马汀马志中TheeffectsofvariedantigenretrievalmethodsonimmunohistochemistryoffunctionalproteinsinretinalparaffinsectionLiYing,HuYuntao,MaTing,MaZhizhong.PekingUniversityThirdHospital,PekingU
2、niversityEyeCenter,Beijing100191,ChinaAbstractBackgroundAntigenretrievalmethodisthekeyofimprovingthesuccessfulrateofimmunohistochemicalassayinparaffinsections.Tostudyanavailablemethodofantigenretrievalisagoaltoachievebothgoodimmunochemistryresult
3、andpreservingretinalproteins.ObjectiveTheaimofpresentstudyistoinvestigatetyrosindigestion,hightemperatureheatpressure,waterbathheatingandmicrowaverepairinantigenretrievalforretinaltissues.MethodsRetinaltissuewasisolatedandobtainedfromcleanChinch
4、illarabbits.Fourhundredsretinalparaffinsectionswereprepared.Fourkindsofantigenretrievalmethodsforretinaltissueincludingtyrosindigestion,hightemperatureheatpressure,waterbathheatingandmicrowaverepairwereusedrespectively.Thedepigmentatedretinalparaffi
5、nsectionwithoutantigenretrievalwasusedascontro.lThepositiveratesofexpressionofCRALBP,Rhodopsinandopsinproteinswereevaluatedandcomparedamongfourkindsofantigenretrievalmethodsbyimmunochemistry.ResultsCRALBP,Rhodopsinandopsinproteinwaspositivelyexpres
6、sedincytoplasmofretinalpigmentepithelialcellsandtheoutersegmentofphotoreceptorrespectively.Nosignificantdifferencewasfoundinthepositiveexpressionratesofthesethreeproteinsamongfourkindsofantigenretrievalmethods(P>005),butthedifferencesintissueintegri
7、tyandbackgroundstainingwerestatisticallysignificant(P<001).Thestructuraldamageofretinaincludedlossandpuckerofscalera,crackofnucleusandabnormalbackgroundstaininhightemperatureheatpressuremethod,waterbathheatingmethodandmicrowaveretrievalmethod.Howev
8、er,stableCRALBP,Rhodopsinandopsinproteinexpressionandstraineffectiveness,clearbackgroundwithoutunspecificstainingandintegratedtissuewereseenintyrosindigestionmethod.ConclusionDuringtheclinicalpathologyanalysisofretinaltissue,theapplicationoftyrosin
