PrinciplesofFluorescenceSpectroscopy.pptVIP

Principles of Fluorescence Spectroscopy Chemistry Department XMU Introduction to Fluorescence 1.0 Introduction 1.1 Phenomenon of Fluorescence 1.2 Excitation and Deactivation of molecule 1.3 Models of Molecular emission 1.4 Characteristics of Fluorescence Emission 1.5 Characteristics of Fluorophore 1.6 Fluorescence quenching 1.7 Resonance energy transfer 1.8 Time scale of Fluorescence 1.9 Intensity and Concentration 1.10 Fluorophore 1.5 Characteristics of fluorophore 1.5.1 Excitation wavelength 激发波长 ?ex 1.5.2 Emission wavelength 发射波长 ?em 1.5.3 Extinction (absorption) coefficient 吸光系数 ? 1.5.4 Stokes’ shift 1.5.5 Fluorescence lifetime 荧光寿命 ? 1.5.6 Fluorescence quantum yield 荧光量子产率 ? 1.5.7 Fluorescence Anisotropy 各项异性 r 15.8 Fluorescence Polarization 荧光偏振 p Absorption and emission spectra Excitation and emission spectra Three dimension spectrum Three dimension spectra 1.5.5 Fluorescence lifetime Expression 1.5.6 fluorescence quantum yield Determination Choose Standards 1.5.7 Fluorescence Anisotropy 荧光分子 Principle of photoselective excitation Definition Polarization and anisotropy 各项异性的物理意义 Isotropic solution 坐标系中的荧光体 Magic angle 魔鬼角 激发跃迁距相对于发射跃迁距取向 Losing of anisotropy 1.6 Fluorescence quenching 1.7 Fluorescence resonance energy transfer 1.7 Fluorescence resonance energy transfer 1.8 Time scale of molecular process in solution Probing pH Probing polarity 1.8 Time scale of molecular process in solution 1.8 Time scale of molecular process in solution Structure of DPPG 1.9 Intensity and concentration 1.10 Intrinsic fluorophore and extrinsic fluorophore Protein fluorescence Protein fluorescence membrane membrane DNA Cofactor 辅酶 Other fluorophores Fluorescent indicators F Ag F-Ag Ab F-Ag:Ab 发射消偏振 旋转快 旋转慢 保持偏振发射 垂直偏振激发 e.g. Rotation diffusion of protein during the lifetime of fluorophores e.g. Rotation diffusion of fluorophore in the excited-state lifetime, probing properties of membrane. Emi