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1、 Histology Chapter1:IntroductionofHistology1. What is Histology? 2. Why to study Histology? 3. What methods are used to study Histology? What is Histology ?Histology：a subject to study themicrostructureofnormalhuman body and itsrelatedfunction.Microanatomy1. Concept of Histology2.StudyingcontentsofH
2、istology：withdifferentshape,sizeandfunction(1)Cell: thesmalleststructuralandfunctionalunit. 1). Cell membrane：unitmembrane Its mainly composed ofa lipid bilayer of phospholipid molecules but with a large numbersof protein molecules protruding the layer. 2).Cytoplasm：OrganellesRER,ribosome-proteinSER
3、-glycogen,lipid,detoxicationMitochondriaGolgicomplex,lysosome,microbody(peroxisome)microfilament,microtubule,intermediatefilament,centrosome3).Nucleus：chromatinchromosomes-DNA,genes（2）Tissue：1.Concept:Thecellswithsimilarshapeandfunctionintercellularsubstance（extracellularmatrix)2.Classificationofthe
4、tissue(primary ,fundamental or basic tissues) 1. Epithelial tissue 2. Connective tissue 3. Muscle tissue 4. Nervous tissue General Histology(3) Organ: Arelativelyindependentpartofthebodythatcarriesoutoneormorespecialfunctions.Examplesoforgansincludetheeyes,ears,heart,lungs,andliver.(4) System：agroup
5、ofbodyorgansorstructuresthattogetherperformoneormorevitalfunctionslikecirculatorysystem,digestivesystem,endocrinesystem.SpecialHistologyCell tissue organ systemWhy to study Histology ?vTocompletetheknowledgeofhumanbodysstructures-fromgrosstomicroscopicvBeabletounderstandhowthedifferenttissuesfunctio
6、n-thebasisofphysiologyvCanfindthediseasesonlyafterthenormalisknown-thebasisofpathologyvItisrelatedtosomemodernsciencefields:cellapoptosis,cellrecognition,embryostemcells,eugenicsandetc.vItisalsoafoundationofclinicsciencesforagooddoctorneededinfuturelBecauseallknowledgeisworthwhile,and/orbecauselearn
7、ingaboutthetissuecompositionofthehumanbodyissheerjoy.lBecausetheperspectiveofhistologycanilluminatemanyotheraspectsofmedicine,enrichingandstrengthingunderstandinginotherareas.Becausesomeofyourpatientsmightexpectclear,accurateexplanationsofhowandwhytheircellsaremisbehaving.lBecausesomedayyoumightbeco
8、meapathologist,andhistologyinterpretationisanessentialclinicalskillforapathologist.lBecausesomedayyoumightneedtocommunicatewithapathologist.lBecauseyouwanttoscorewellontheU.S.MedicalLicensingExam.Unit used in microscope 1m=1/1000mm 1nm=1/1000mMaximum resolutionLightmicroscope:0.2mTransmissionelectro
9、nmicroscope:0.2nm Scanning electronmicroscope:5nmInvestigative methods of HistologyL Light microscopylBaselPillarlStandlStage lCyclinderlMirrorlCondenserlObjectiveslEyepiecelCoarse adjustment knoblFine adjustment knob The process of the routine HE slide preparation:1.Excisionandfixation：fixatives:Al
10、dehydes,Alcohols,Picrates coagulate protein2.Dehydratingandembedding：ascendinggradesofalcoholclearwithxyleneorchloroform(organicsolvent)embedthetissueinparaffin3. Sectioning ：5-8mmicrotomemount on slides deparaffin hydrating stained by H&E methodother preparations:Cryostat:frozensectionVibratomeSmea
11、rWholeMountsGroundSectionH-E Staining:lHematoxylin: basic dye, purple-bluelEosin: acid dye, pink colorlBasophilic: components bonded by basic dye (H); purple-blue（nuclear chromatin & basophilic substance in cytoplasm）l Acidophilic: components bonded with acid dye (E); pink (cytoplasm & collagenous f
12、iber)lNeutrophilic:donotstainw/neitherbasicnoraciddyesMetachromasia:adyestainstissueadifferentcolorfromthatofthedyesolutionitself,e.g.toluidinebluestainsmastcellsinpurplecolorSpecialstaining:ArgyrophiliaArgentaffinSilverstainingofSilverstainingoftheneuronandtheneuronandthebilecanaliculithebilecanali
13、culiToluidinebluestainingofmastcells4.Clearingandenveloping：xylinCanadabalsamcover-glass1.TransmissionElectronMicroscope(TEM)1.TransmissionElectronMicroscope(TEM) Usingabeamofelectrons(shortwave-lengths)Usingabeamofelectrons(shortwave-lengths)insteadofvisiblelight.insteadofvisiblelight.SSectionectio
14、n preparation:preparation:similartothoseforL.Mmainly,similartothoseforL.Mmainly,plasticinsteadofparaffin,plasticinsteadofparaffin,50-70nmthick(ultrathinsection),50-70nmthick(ultrathinsection),heavymetalsaltsinsteadofHE.heavymetalsaltsinsteadofHE. ResolutionResolution:0.1-0.5:0.1-0.5nm nm (0.2nm)(0.2
15、nm) Ultrastructure:Ultrastructure:ThestructureovservedinEMThestructureovservedinEM Electron-dense(moreElectron-dense(more structuresstructures stainedstained withwith heavyheavy matalmatal salts)salts) / /electron-electron-lucentlucent(lessstructuresstainedwithheavymetalsalts)(lessstructuresstainedw
16、ithheavymetalsalts)II.ElectronMicroscopyTransmission Electron MicroscopeDiagramsofTEM2.ScanningElectronMicroscope(SEM)Showingthe3dimensionalsurfaceArchitectureofcellsandtissuesResolution:5nmIII.Histochemistry & CytochemistryRevealthechemicalcompositioninsitu(e.g.proteins,a.a.,nucleicacid,lipids,enzy
17、mesetc.)w/chemical,biochemicalmethods. Theproductofchemicalreactionshouldbeinsoluble/colored/electron-scattering,&beseeninLMorEM polysaccharide + HIO4 (periodic acid) (1,2-glycolgroup)(oxidise)Aldehyde group + Shiffs reagent (colorless) Purplish red depositorFor instance:vPAS( (P PeriodicAcidSchiff)
18、reaction: formanifestingpolysaccharideandproteoglycan(e.g.glycogen). PASreactioninthehepatocytesGobletcellsvImmunocytochemistryBasedonantigenbindstospecificantibody. Tissuesectionw/Antigen+ labeled antibodylabeledAg-AbcomplexFluoresceinlabellingenzymelabellingcolloidalgoldcolloidalgoldlabellinglabel
19、lingNOS positive neuron in hypothalamus of ratNOS & GnRH positive NeuronsNOS & GnRH positive Neurons in hypothalamus of rat in hypothalamus of ratThe methodsinlearning H HistologyCombinationofthe2dimentionalstructurewith3dimentionCombinationofthetheorywithpracticeCombinationofthestructurewithfunctionConcernthedynamicchange
