顯微鏡種類與背景介紹教學(xué)課件

Scale1Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.DiscoveryofMicroorganismsAntonyvanLeeuwenhoek(1632-1723)firstpersontoobserveanddescribemicro-organismsaccuratelyFigure1.1b2Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.LensesandtheBendingofLightlightisrefracted(bent)whenpassingfromonemediumtoanotherrefractiveindexameasureofhowgreatlyasubstanceslowsthevelocityoflightdirectionandmagnitudeofbendingisdeterminedbytherefractiveindexesofthetwomediaformingtheinterface3Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Lensesfocuslightraysataspecificplacecalledthefocalpointdistancebetweencenteroflensandfocalpointisthefocallengthstrengthoflensrelatedtofocallengthshortfocallengthmoremagnification4Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.25Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.TheLightMicroscopemanytypesbright-fieldmicroscopedark-fieldmicroscopephase-contrastmicroscopefluorescencemicroscopesarecompoundmicroscopesimageformedbyactionof2lenses6Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.TheBright-FieldMicroscopeproducesadarkimageagainstabrighterbackgroundhasseveralobjectivelensesparfocalmicroscopesremaininfocuswhenobjectivesarechangedtotalmagnificationproductofthemagnificationsoftheocularlensandtheobjectivelens7Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.38Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.49Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.MicroscopeResolutionabilityofalenstoseparateordistinguishsmallobjectsthatareclosetogetherwavelengthoflightusedismajorfactorinresolutionshorterwavelengthgreaterresolution10Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.workingdistance

distancebetweenthefrontsurfaceoflensandsurfaceofcoverglassorspecimen11Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.512Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.613Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.TheDark-FieldMicroscopeproducesabrightimageoftheobjectagainstadarkbackgroundusedtoobserveliving,unstainedpreparations14Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.7b15Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.ThePhase-ContrastMicroscopeenhancesthecontrastbetweenintracellularstructureshavingslightdifferencesinrefractiveindexexcellentwaytoobservelivingcells16Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.917Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.1018Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.TheDifferentialInterferenceContrastMicroscopecreatesimagebydetectingdifferencesinrefractiveindicesandthicknessofdifferentpartsofspecimenexcellentwaytoobservelivingcells19Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.TheFluorescenceMicroscopeexposesspecimentoultraviolet,violet,orbluelightspecimensusuallystainedwithfluorochromesshowsabrightimageoftheobjectresultingfromthefluorescentlightemittedbythespecimen20Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.1221Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.13candd22Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.PreparationandStainingofSpecimensincreasesvisibilityofspecimenaccentuatesspecificmorphologicalfeaturespreservesspecimens23Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Fixationprocessbywhichinternalandexternalstructuresarepreservedandfixedinpositionprocessbywhichorganismiskilledandfirmlyattachedtomicroscopeslideheatfixingpreservesoverallmorphologybutnotinternalstructureschemicalfixingprotectsfinecellularsubstructureandmorphologyoflarger,moredelicateorganisms24Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.DyesandSimpleStainingdyesmakeinternalandexternalstructuresofcellmorevisiblebyincreasingcontrastwithbackgroundhavetwocommonfeatureschromophoregroupschemicalgroupswithconjugateddoublebondsgivedyeitscolorabilitytobindcells25Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.DyesandSimpleStainingsimplestainingasinglestainingagentisusedbasicdyesarefrequentlyuseddyeswithpositivechargese.g.,crystalviolet26Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.DifferentialStainingdividesmicroorganismsintogroupsbasedontheirstainingpropertiese.g.,Gramstaine.g.,acid-faststain27Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.GramstainingmostwidelyuseddifferentialstainingproceduredividesBacteriaintotwogroupsbasedondifferencesincellwallstructure28Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.14primarystainmordantcounterstaindecolorizationpositivenegative29Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.15cEscherichiacoli–agram-negativerod30Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Acid-faststainingparticularlyusefulforstainingmembersofthegenusMycobacteriume.g.,Mycobacteriumtuberculosis–causestuberculosise.g.,Mycobacteriumleprae–causesleprosyhighlipidcontentincellwallsisresponsiblefortheirstainingcharacteristics31Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.StainingSpecificStructuresNegativestainingoftenusedtovisualizecapsulessurroundingbacteriacapsulesarecolorlessagainstastainedbackground32Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.StainingSpecificStructuresSporestainingdoublestainingtechniquebacterialendosporeisonecolorandvegetativecellisadifferentcolorFlagellastainingmordantappliedtoincreasethicknessofflagella33Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.ElectronMicroscopybeamsofelectronsareusedtoproduceimageswavelengthofelectronbeamismuchshorterthanlight,resultinginmuchhigherresolutionFigure2.2034Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.TheTransmissionElectronMicroscopeelectronsscatterwhentheypassthroughthinsectionsofaspecimentransmittedelectrons(thosethatdonotscatter)areusedtoproduceimagedenserregionsinspecimen,scattermoreelectronsandappeardarker35Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.23EM36Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.SpecimenPreparationanalogoustoproceduresusedforlightmicroscopyfortransmissionelectronmicroscopy,specimensmustbecutverythinspecimensarechemicallyfixedandstainedwithelectrondensematerial37Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Otherpreparationmethodsshadowingcoatingspecimenwithathinfilmofaheavymetalfreeze-etchingfreezespecimenthenfracturealonglinesofgreatestweakness(e.g.,membranes)38Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.2539Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Ebola40Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Flyhead41Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.TheScanningElectronMicroscopeuseselectronsreflectedfromthesurfaceofaspecimentocreateimageproducesa3-dimensionalimageofspecimen’ssurfacefeatures42Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.Figure2.2743Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.NewerTechniquesinMicroscopyconfocalmicroscopyandscanningprobemicroscopyhaveextremelyhighresolutioncanbeusedtoobserveindividualatomsFigure2.2044Copyright?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.?TheMcGraw-HillCompanies,Inc.Permissionrequiredforreproductionordisplay.ConfocalMicroscopyconfocalscanninglasermicroscopelaserbeamusedtoilluminatespotsonspecimencomputercompilesimagescreatedfromeachpointtogeneratea3-dimensionalimage45Copyright?TheMcGr