简介：ObjectivesTodevelopasimple,accurateandreproduciblemethod,whichcombinesmacroandhistopathologicaltechniquesfordeterminingthedegreeoflipiddepositioningeneticallymodifiedmice.MethodTheentireaortasfromC57BL/6,ldlr-/-andapoE-/-micewerestainedwithSudanⅣusingeitherinvivoperfusionortraditionalinvitroenfacestainingtechniques.Histologicalsectionsofaorticrootandheartswereembeddedintissuefreezingmediumandcutwithacryostat,thenstainedwithOilRedO.Thecalculatedaorticrootareabasedontheaorticrootcircumferencewasusedtoreducemeasurementerrors.ResultsTheinvitroenfacestainingcanstainallfat,whichincludetheadventitialtissuearoundaorta.Howevertheinvivoperfusionstainingcanspecificallystainthefattydepositioninsideofaorta.Bothentireaortaandaorticrootsectionstainingshowedthattherewasahighlysignificantincreaseinfattydepositionintheaortasofthegeneticmodifiedmice.Althoughallmicegeneticbackgroundwassame,theapoE-/-micehadlargeratheroscleroticlesionsthanldlr-/-mice.ConclusionsThenewinvivoperfusionmethodismoreaccuratethantheinvitroenfacemethod.Thecombinationofthesemacroandmicroscopictechniquesovercomestheshortcomingsoftheearlierpublishedmethodswhicharegenerallylimitedtothemeasurementoffattyredstainingareasonly,neglectingnon-specificadventitialfatstainingaroundaortaandaorticrootsectiontissuedistortion.
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