BriefreportSimultaneousdetectionofacellsurfaceantigenandapoptosisbymicrowave-sensitizedTUNELassayonparaffinsectionsMayerlChristinaa,Hammerer-LercherAngelikab,PuschendorfBerndb,MartinaPrelogc,⁎aDivisionofExperimentalPathophysiologyandImmunology,BiocenterInnsbruck,InnsbruckMedicalUniversity,AustriabDivisionofClinicalBiochemistry,BiocenterInnsbruck,InnsbruckMedicalUniversity,AustriacDepartmentofPediatrics,InnsbruckMedicalUniversity,Anichstr.35,A-6020Innsbruck,AustriaReceived13December2005;receivedinrevisedform9August2006;accepted10August2006Availableonline20September2006AbstractTheTUNEL(terminaldeoxynucleotidyltransferase-mediateddUTPnick-endlabeling)techniquehasbeendescribedasasensitivemethodfordetectionofapoptoticnucleiintissuesandpreferentialstainingofapoptoticstrandbreaks.Short-termmicrowavepre-treatment,anon-enzymaticpre-treatmenttechniqueofantigenretrieval,hasbeendemonstratedtooptimizetheTUNELmethodforinsitudetectionofapoptoticcellsinformalin-fixedparaffin-embeddedtissuesections.Inthepresentstudy,wesensitizedinternalmammaryarterysectionsbyshort-termmicrowavepre-treatmentandusedatwo-stepindirectenzymaticmethodtogainasanendproductdifferentiallystainedcells,namelyTUNEL-positivecellsandthesepositiveforthesurfacemarkervonWillebrandfactor(vWF).Thistechniqueenablestoclearlydistinguishbetweenapoptotic,non-apoptoticandvWF-positivecellsthatarephenotypicforendothelialcells.Phenotypicidentificationofcellsissimplifiedbydoublestainingwithcellsurfacemarkers.Thisrapid,sensitiveandreproducibletechniqueallowssimultaneousdetectionofDNAfragmentationandphenotypicmarkersinthesameparaffin-embeddedhumantissuesection.©2006ElsevierB.V.Allrightsreserved.Keywords:TUNEL;Apoptosis;vonWillebrandfactor;Doublestaining;Microwaveirradiation1.IntroductionSinceGavrielidescribedtheterminaldeoxynucleoti-dyltransferase(TdT)mediateddeoxyuridinetriphosphate(dUTP)nickendlabeling(TUNEL)techniquein1992(Gavrielietal.,1992),ithasbeenextensivelyappliedontissuesectionsaswellascellculturesforidentificationo...
