One-Step Preservation of Phosphoproteins and Tissue Morphology at Room Temperature for Diagnostic and Research Specimens 英文参考文献.docVIP

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One-Step Preservation of Phosphoproteins and Tissue Morphology at Room Temperature for Diagnostic and Research Specimens 英文参考文献.doc

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One-Step Preservation of Phosphoproteins and Tissue Morphology at Room Temperature for Diagnostic and Research Specimens 英文参考文献

One-StepPreservationofPhosphoproteinsandTissue MorphologyatRoomTemperatureforDiagnosticand ResearchSpecimens ClaudiusMueller1*,KirstenH.Edmiston2,CalvinCarpenter3,EoinGaffney4,CiaraRyan4,RonanWard4, SusanWhite4,LorenzoMemeo5,CristinaColarossi5,EmanuelF.Petricoin,III1,LanceA.Liotta1 ,Virginia Espina1 1CenterforAppliedProteomicsandMolecularMedicine,GeorgeMasonUniversity,Manassas,Virginia,UnitedStatesofAmerica,2CancerTranslationalResearchCenter, InovaFairfaxHospital,InovaHealthSystem,FallsChurch,Virginia,UnitedStatesofAmerica,3CenterforBiodefenseandInfectiousDisease,GeorgeMasonUniversity, Manassas,Virginia,UnitedStatesofAmerica,4St.James’sHospital,Dublin,Ireland,5PathologyUnit,MediterraneanInstituteofOncology,Catania,Italy Abstract Background: There is an urgent need to measure phosphorylated cell signaling proteins in cancer tissue for the individualization of molecular targeted kinase inhibitor therapy. However, phosphoproteins fluctuate rapidly following tissue procurement. Snap-freezing preserves phosphoproteins, but is unavailable in most clinics and compromises diagnosticmorphology.Formalinfixationpreservestissuehistomorphology,butpenetratestissueslowly,andisunsuitable forstabilizingphosphoproteins.Weoriginatedandevaluatedanovelone-stepbiomarkerandhistologypreservative(BHP) chemistrythatstabilizessignalingproteinphosphorylationandretainsformalin-liketissuehistomorphologywithequivalent immunohistochemistryinasingleparaffinblock. Results:TotalproteinyieldextractedfromBHP-fixed,routineparaffin-embeddedmouseliverwas100%comparedtosnap- frozentissue.Theabundanceof14phosphorylatedproteinswasfoundtobestableoverextendedfixationtimesinBHP fixedparaffinembeddedhumancolonmucosa.Comparedtomatchedsnap-frozentissue,8phosphoproteinswereequally preservedinmouseliver,whileAMPKb1Ser108wasslightlyelevatedafterBHPfixation.Morethan25tissuesfrommouse, catandhumanspecimenswereevaluatedforpreservationofhistomorphology.Selectedtissueswereevaluatedinamulti- site,independentpathologyrevie