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1、1現(xiàn)代分子熒光檢測(cè)技術(shù)及應(yīng)用王蘭芬博士市場(chǎng)部分子光譜工程師18/02/20142熒光現(xiàn)象的發(fā)現(xiàn)?1845年,英國(guó)的弗雷德里希等人首次觀察到奎寧的藍(lán)色450nm的發(fā)光(熒光).YellowglassofwineEmfilter>400nmBlueglassQuinineFilter<400nmSolution3發(fā)光類型PhenomenonMdfModeofexcitittiationPhotoluminescence(fluorescence,Absorptionoflight(photons)phosphorescence)(光致發(fā)光)Radioluminescence(輻射發(fā)光)Io
2、nizingradiation(X-rays?????????)Cathodoluminescence(陽極輻射發(fā)光)Cathoderays(electronbeams)Electroluminescence(電致發(fā)光)ElectricfieldThermoluminescence(熱致發(fā)光)Heatingafterpriorstorageofenergy(e.g.radioactiveirraditidiation)Chemiluminescence(化學(xué)發(fā)光)Chemicalprocess(e.g.oxidation)Bioluminescence(生物發(fā)光)Biochemical
3、processTriboluminescence(摩擦發(fā)光)FrictionalandelectrostaticforcesSonoluminescence(超聲發(fā)光)Ultrasounds4熒光的發(fā)展史?1853(GGStokesG.G.Stokes)提出熒光概念?1858(E.Becquerel)首次觀察到磷光?1867(F.Goppelsr?der)熒光分析鋁-桑色素復(fù)合物測(cè)定鋁離子?1888(EE.WidWiedemann)提出發(fā)光概念?1926(F.Perrin)熒光偏振理論?1926(Gaviola)第一臺(tái)熒光壽命測(cè)量?jī)x器(相調(diào)制原理)?1935(A.Jablonski):
4、雅布隆斯基能級(jí)圖(Jablonski’sdiagram)?…5熒光的產(chǎn)生機(jī)理AbsorptionrsioneSexcitedstate2ylConvgaInternenerSexcitdttitedstate1FluorescenceNonradiativedissipationGroundStateGroundStateElectrons6磷光的產(chǎn)生機(jī)理AbsorptionergyS2ICceennsorbabAIntersystemCrossingS1T1PhosphorescenceGroundStateElectrons7JablonskiJablonskiDiagramDia
5、gram2S120A=PhotonabsorptionF=FlouresceneICT=PhosphoresceneS1S=ElectrongroupstateT02S=ElectronfirstVR1ISC210ICexcitationsingletTstate1VRS=ElectronsecondEnergy2excitdiitedsinglltttetstateT=Electronfirst1AAFTexcitedtripletstateT=Electronsecond=Electronsecond2excitationtripletstate2IC=Internalconver
6、sionIC=InternalconversionS1ISC=Intersystem00Absorption:10-15SconversioInternalconversionandvibrationalrelaxation:10Internalconversionandvibrationalrelaxation:10VRVRvibration-14=-vibration10-11SrelaxtionFluorescene:10-9-10-6S,nsorderPhosphorescene:10-3-10-2S8熒光技術(shù)中的幾個(gè)基本概念?熒光發(fā)射譜、激發(fā)譜、三維熒光譜量子產(chǎn)率熒光壽命:?
7、斯托克斯位移(StokesshifStokesshift)?同步譜?三維光譜?內(nèi)濾效應(yīng)(Innerfiltereffect)?熒光各向異性和偏振(Anisotropyandpolarization):randP?熒光淬滅FRETFRAP9熒光發(fā)射譜和激發(fā)譜ytexcitationemissionensitInWlhWavelengthexcitation:固定發(fā)射波長(zhǎng),激發(fā)波長(zhǎng)掃描,觀察物質(zhì)發(fā)光特性的激發(fā)光依賴性emission:固定激發(fā)波長(zhǎng),