Tissue Harvest Protocol

TISSUESTOBEPROCURED(minimallyandpreferablywithin5-8hoursafterdeath):1.Brain2.Liver3.Muscle4.Skin5:Othersasthespecificcasedictates PROCESSINGANDSTORAGEOFTISSUES:1.Brain Removethebrain,withcerebellumandbrainstemintact;cutmid-sagitallyandseparatethecerebralhemispheres. Fix1/2ofbraininformalinandstoreforseveralweeks;thendrainoffexcessformalinandrepackageinaheat-sealedplasticbag(x2)orairtightplasticcontainerandFEDEXtotherepository,billingrecipient(seenotes). Removethebrainstemandcerebellumfromtheotherhalfofthebrainbysectioningthroughthecerebralpeduncles. Sectionthecerebellum,longitudinally,justlateraltothemostlateralextentofthebrainstem. Sectiontheremaininghemisphere,placingthemedialsurfacedown,coronallyat1cmintervalsfromfrontaltooccipitalpoles. Withtheremainderofthebrain:placeeachsectiononafreezingtray(aluminumfoiloverablockofdryiceorifablockisunavailable,aluminumfoiloverastainlesssteeltrayoverpelletsofdryice).Coverthesectionswithpowdereddryiceastheyareplaced. Oncethesectionsarefrozen(about5minutes),wrapeachinthefoilandlabelintheordereachsectionwascut.Thesefoil-wrappedsectionscanbeplacedinaziplocfreezerstoragebagandstoredinthe-70°Cfreezeruntiltheyareshippedtotherepository(seenotes). 2.LiverPlaceapproximately1gramofliverintothesamecontaineroffixativeasthebrain.Freezea1cmthickslabofliverinthesamemannerasthebrain. 3.MusclePlaceapproximately1gofmuscleintothesamefixativecontainerasthebrain.Freezeapproximately10gofmuscleinthesamemannerasthebrain(generally,anyskeletalmusclewilldobutitmaydependontheclinicalhistory). 4.SkinPlacea10mmx3mmpieceofskinwithunderlyingADIposeremovedintosterileculturemediuminasterilefalcontubeandFEDEXtotherepository.Ifafibroblastcultureexists,thisstepwillnotbenecessary.

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