组织学与胚胎学绪论中英课件

HistologyIntroduction YuXiangmin(于向民）DepartmentofHistologyandEmbryology1.HistologyYuXiangmDefinition:A science:study normal micro-structure(微微细结构构)&its related function of human body.4 structural levels:Cell:thesmalleststructural&functionalunit.Tissue(组织):groups ofcells(similarinmorphologyorrelatedinfunction)intercellular materials4typesoffundamentaltissueepithelium上皮上皮组织connectivetissue结缔组织musculartissue肌肌组织nervoustissue神神经组织2.Definition:Ascience:studyn Organs:器官organizationsofvariouskindsoftissuesinparticularways&performaspecific function.System:系统formedbyseveralfunction-relatedorganswhichtogetherperformacontinuousphysiologicalfunction.Forexample:digestivesystem3.Organs:器官organizationso4.4.Why do we study histology?vTocompletetheknowledgeofhumanbodysstructures-fromgrosstomicroscopicvBeabletounderstandhowthedifferenttissuesfunction-thebasisofphysiologyvCanfindthediseasesonlyafterthenormalisknown-thebasisofpathologyvItisalsoafoundationofclinicsciencesforagooddoctorneededinfuture5.Whydowestudyhistology?ToInvestigative methods of histologyI.Light microscopy6.Investigativemethodsofhistol1.Tissue preparation A.A.paraffin section preparation石蜡切片石蜡切片石蜡切片石蜡切片术术v Specimen取材取材取材取材:asfreshaspossibleFixation固定固定固定固定:fixative:formalinsolution;purpose:topreservethestructuralorganization Dehydration脱脱脱脱水水水水:replacethewaterinthetissuebyalcoholClearing透明透明透明透明:replacethealcoholbyxyleneEmbedding包埋包埋包埋包埋:replacethexylenewithmeltedparaffinSectioning&mounting切片切片切片切片7.l1.Tis8.8.B.Frozen section:Betterforpreservingchemicalcomponents(e.g.enzymes)FreezingcryostatstainingC.the others:Smearpreparations:forbloodetc;Grindpreparations:forbone9.B.Frozensection:9.2.Staining染色染色v Purpose:Tomaketissuesectionforobservation.vH-E Staining:Hematoxylin:苏木精木精 basic dye,bluishcolorEosin:伊伊红aciddye,pinkcolor Basophilic:嗜碱性嗜碱性componentsstainedwithbasicdye(H);bluish（nuclearchromatin)Acidophilic:嗜酸性嗜酸性componentswithanaffinityforacidicdye(E);pink(cytoplasm&collagenfiber)10.2.Staining染色10.Neutrophilic:中中 性性donotstainwithbothbasicandaciddyesArgyrophilia:嗜嗜银性性 thosewithanaffinityforsilver,dark-browncolor 11.Neutrophilic:中性11.FluorescentstainingshowstheapoptosisinhumanHL-60cellsSilverstainingofSilverstainingoftheneuronandtheneuronandthebilecanaliculithebilecanaliculi12.Fluorescentstainingshowsthe1.1.Transmission Electron Microscope (TEM)Transmission Electron Microscope (TEM)透射透射透射透射电镜电镜Usingabeamofelectrons(shortwave-lengths)Usingabeamofelectrons(shortwave-lengths)insteadofvisiblelight.insteadofvisiblelight.Sectionpreparation:Sectionpreparation:similartothoseforL.Mmainly,similartothoseforL.Mmainly,plasticinsteadofparaffin,plasticinsteadofparaffin,50-70nmthick,50-70nmthick,heavymetalsaltsinsteadofHE.heavymetalsaltsinsteadofHE.ResolutionResolution:0.1-0.5:0.1-0.5nm nm Ultrastructrue:Ultrastructrue:超微超微超微超微结结构构构构ThestructureinEMThestructureinEM Electron-denseElectron-dense电电子密度高子密度高子密度高子密度高 electron-lucentelectron-lucent电电子密度低子密度低子密度低子密度低II.Electron Microscopy13.TransmissionElectronMicroscoTransmission Electron Microscope14.TransmissionElectronMicrosco15.15.DiagramsofTEM16.DiagramsofTEM16.2.Scanning Electron Microscope(SEM)2.Scanning Electron Microscope(SEM)扫扫描描描描电镜电镜Sowingthe3dimensionalsurfaceArchitectureofcellsandtissuesResolution:5nm17.2.ScanningElectronMicroscoJEOL扫描描电子子显微微镜18.JEOL扫描电子显微镜18.III.Histochemistry&Cytochemistry 组织化学化学术Revealthechemicalcompositioninsitu(e.g.proteins,a.a.,nucleicacid,lipids,enzymesetc.)withchemical,biochemicalmethods.vTheproductofchemicalreactionshouldbeinsoluble/colored/electron-scattering,&beseeninLMorEM 19.III.Histochemistry&Cytochempolysaccharide +HIO4 (hydroxylgroup)(oxidise)Aldehyde group +Shiffs reagent(colorless)Purplish red depositorFor instance:vPAS(P(PeriodicAcidSchiff)reaction:formanifestingpolysaccharide多糖多糖 (糖原）糖原）20.polysaccharide+HIO4ForinPAS reaction in the goblet cell21.PASreactioninthegobletcelIV.Immunocytochemistry免疫免疫组织化学化学Basedonantigenbindstospecificantibody.TissuesectionwithAntigen+labelled antibodylabelledAg-AbcomplexFluoresceinlabellingenzymelabellingcolloidalgoldcolloidalgoldlabellinglabelling22.IV.Immunocytochemistry免疫组织化学23.23.V.tissue culture 组织培养培养VI.in situ hybridization(ISH)nucleic acid molecular hybridization 原位原位杂交交术 Use nucleotide probe to check targetfragment of intracellular DNA or mRNA insitu,in order to study the gene expression.24.V.tissueculture组织培养ExpressionofPSAandPSAmRNAinhumanprostate(histochemistry&ISH)25.ExpressionofPSAandPSAmRNThe methodinlearning histologyCombinationofthe2dimentionalstructurewith3dimentional26.ThemethodinlearninghistoloCombination of the theory with practiceCombination of the structure with functionConcern the dynamic change27.CombinationofthetheorywithQuestionsforreviewvWhatistheH&Estaining?vDescribebasophilic,acidophilic,neutrophilic?vWhatdoesPASpositivemeans?28.QuestionsforreviewWhatisth