組織學(xué)與胚胎學(xué)緒論中英

Histology

Introduction

YuXiangmin(于向民）DepartmentofHistologyandEmbryologyDefinition:

Ascience:studynormalmicro-structure(微細(xì)結(jié)構(gòu))&itsrelatedfunctionofhumanbody.

4structurallevels:

Cell:thesmalleststructural&functionalunit.

Tissue(組織):

groupsof

cells(similarinmorphologyorrelatedinfunction)＋intercellularmaterials

4typesoffundamentaltissue

epithelium上皮組織

connectivetissue結(jié)締組織

musculartissue肌組織

nervoustissue神經(jīng)組織

Organs:器官organizationsof

variouskindsoftissuesinparticularways&performaspecific

function.

System:系統(tǒng)formedbyseveralfunction-relatedorganswhichtogetherperformacontinuousphysiologicalfunction.Forexample:digestivesystem

Whydowestudyhistology?Tocompletetheknowledgeofhumanbody’sstructures----fromgrosstomicroscopicBeabletounderstandhowthedifferenttissuesfunction----thebasisofphysiologyCanfindthediseasesonlyafterthenormalisknown----thebasisofpathologyItisalsoafoundationofclinicsciences—foragooddoctorneededinfutureInvestigativemethodsofhistologyI.Lightmicroscopy

l

1.Tissuepreparation

A.paraffinsectionpreparation石蠟切片術(shù)

Specimen取材:

asfreshaspossible

Fixation固定:fixative:formalinsolution;purpose:topreservethestructuralorganization

Dehydration脫水:replacethewaterinthetissuebyalcohol

Clearing透明:replacethealcoholbyxylene

Embedding包埋:replacethexylenewithmeltedparaffin

Sectioning&mounting切片B.

Frozensection:

Betterforpreservingchemicalcomponents(e.g.enzymes)Freezing→cryostat→stainingC.theothers:Smearpreparations:forbloodetc;Grindpreparations:forbone

2.Staining染色

Purpose:Tomaketissuesectionforobservation.

H-EStaining:

Hematoxylin:蘇木精basic

dye,

bluishcolor

Eosin:伊紅aciddye,pinkcolor

Basophilic:嗜堿性componentsstainedwithbasicdye(H);bluish（nuclearchromatin)

Acidophilic:嗜酸性componentswithanaffinityforacidicdye(E);pink(cytoplasm&collagenfiber)

Neutrophilic:中性donotstainwithbothbasicandaciddyesArgyrophilia:嗜銀性

thosewithanaffinityforsilver,dark-browncolor

Fluorescentstaining

showstheapoptosisinhumanHL-60cellsSilverstainingoftheneuronandthebilecanaliculiTransmissionElectronMicroscope(TEM)透射電鏡Usingabeamofelectrons(shortwave-lengths)insteadofvisiblelight.Section

preparation:similartothoseforL.Mmainly,plasticinsteadofparaffin,50-70nmthick,heavymetalsaltsinsteadofHE.

Resolution:0.1-0.5nm

Ultrastructrue:超微結(jié)構(gòu)ThestructureinEM

Electron-dense電子密度高

electron-lucent電子密度低ElectronMicroscopy

TransmissionElectronMicroscope

DiagramsofTEM2.ScanningElectronMicroscope(SEM)掃描電鏡Sowingthe3dimensionalsurfaceArchitectureofcellsandtissues

Resolution:5nmJEOL掃描電子顯微鏡III.

Histochemistry&Cytochemistry

組織化學(xué)術(shù)

Revealthechemicalcompositioninsitu(teins,a.a.,nucleicacid,lipids,enzymesetc.)withchemical,biochemicalmethods.Theproductofchemicalreactionshouldbeinsoluble/colored/electron-scattering,&beseeninLMorEM

polysaccharide+HIO4

(hydroxylgroup)(oxidise)Aldehydegroup+Shiff’sreagent

(colorless)

PurplishreddepositorForinstance:PAS(PeriodicAcidSchiff)reaction:

formanifestingpolysaccharide多糖(糖原）PASreactioninthegobletcellIV.Immunocytochemistry免疫組織化學(xué)Basedonantigenbindstospecificantibody.

TissuesectionwithAntigen+

labelled

antibodylabelledAg-AbcomplexFluoresceinlabellingenzymelabellingcolloidalgold

labellingV.tissueculture組織培養(yǎng)VI.insitu

hybridization(ISH)

—nucleicacidmolecularhybridization原位雜交術(shù)UsenucleotideprobetochecktargetfragmentofintracellularDNA

ormRNAinsitu,inordertostudythegeneexpression.ExpressionofPSAandPSAmRNAinhumanprostate(histochemistry&ISH)The

methodinlearning

histologyCombinationofthe2dimentionalstructurewith3diment