荧光偏振及应用
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– θrot is the rotational relaxation (correlation) time (the time required to rotate through an angle whose cosine is 1/e, or approximately 68.5°)
– τfl is the fluorescence lifetime of the excited fluorescent probe
荧光偏振现象
If linear polarized light is used to excite an ensemble of fluorophores, only those fluorophores aligned with the plane of polarization will be excited. There are 2 scenarios for the emission.
r = r0 exp ( –t /θ) Steady-State Fluorescence Polarization Time-Resolved Fluorescence
Jolley and co-workers developed FP into a commercial system for the monitoring of therapeutic drug levels and the detection of drugs of abuse in human body fluids.
荧光偏振现象
荧光偏振&荧光各向异性源自文库
Fluorescence Polarization (FP) P = (I∥ - I⊥) / (I∥ + I⊥)
Fluorescence Anisotropy (FA) r = (I∥ - I⊥) / (I∥ + 2I⊥)
P = 3 r / (2 + r) r = 2 P / (3 – P)
荧光偏振现象
荧光偏振现象
Fluorescence polarization (FP) can be considered a competition between the molecular motion and the lifetime of fluorophores in solution.
荧光偏振及应用
姜浩 2007.3
内容
荧光及其测量 荧光偏振 应用
– 流动性 – 分子基本性质 – 酶学 – 分子相互作用 – 荧光偏振免疫 – 成像
荧光标记方法
荧光
The fluorescence of a molecule is the light emitted spontaneously due to transitions from excited singlet states (usually S1) to various vibrational levels of the electronic ground state, i.e. S1,0→S0,v.
荧光的测量
They carry information about both the photophysical properties of the fluorescing molecule and the chemical and physical nature of its microsurroundings.
荧光的测量
It can be characterized by several parameters.
– Intensity at a given wavelength I(λ)
– Quantum yield
Φ
– Lifetime
τ
– Polarization
P
These parameters can be monitored in a steady-state or time-resolved manner.
荧光
Jabłoński diagram illustrating the creation and fate of a molecular excited singlet state, including absorption (ABS), fluorescence (FL), phosphorescence (PH), internal conversion (IC), intersystem crossing (ISC), vibrational relaxation (VR) and collisional quenching (CQ).Not included are processes like solvent relaxation, energy transfer and photochemical reactions
荧光偏振的测量
荧光偏振的测量
Following a pulse excitation, the fluorescence anisotropy of a spherical particle in a homogeneous isotropic medium decays exponentially
发射光谱 激发光谱 时间分辨 空间分辨
内容
荧光及其测量 荧光偏振 应用
– 流动性 – 分子基本性质 – 酶学 – 分子相互作用 – 荧光偏振免疫 – 成像
荧光标记方法
荧光偏振简史
Fluorescence polarization was first described in 1926 by Perrin.
Weber greatly expanded the theory and developed the first instrumentation for the measurement of FP.
Dandliker and co-workers expanded FP into biological systems, such as antigen-antibody reactions and hormone-receptor interactions.
– τfl is the fluorescence lifetime of the excited fluorescent probe
荧光偏振现象
If linear polarized light is used to excite an ensemble of fluorophores, only those fluorophores aligned with the plane of polarization will be excited. There are 2 scenarios for the emission.
r = r0 exp ( –t /θ) Steady-State Fluorescence Polarization Time-Resolved Fluorescence
Jolley and co-workers developed FP into a commercial system for the monitoring of therapeutic drug levels and the detection of drugs of abuse in human body fluids.
荧光偏振现象
荧光偏振&荧光各向异性源自文库
Fluorescence Polarization (FP) P = (I∥ - I⊥) / (I∥ + I⊥)
Fluorescence Anisotropy (FA) r = (I∥ - I⊥) / (I∥ + 2I⊥)
P = 3 r / (2 + r) r = 2 P / (3 – P)
荧光偏振现象
荧光偏振现象
Fluorescence polarization (FP) can be considered a competition between the molecular motion and the lifetime of fluorophores in solution.
荧光偏振及应用
姜浩 2007.3
内容
荧光及其测量 荧光偏振 应用
– 流动性 – 分子基本性质 – 酶学 – 分子相互作用 – 荧光偏振免疫 – 成像
荧光标记方法
荧光
The fluorescence of a molecule is the light emitted spontaneously due to transitions from excited singlet states (usually S1) to various vibrational levels of the electronic ground state, i.e. S1,0→S0,v.
荧光的测量
They carry information about both the photophysical properties of the fluorescing molecule and the chemical and physical nature of its microsurroundings.
荧光的测量
It can be characterized by several parameters.
– Intensity at a given wavelength I(λ)
– Quantum yield
Φ
– Lifetime
τ
– Polarization
P
These parameters can be monitored in a steady-state or time-resolved manner.
荧光
Jabłoński diagram illustrating the creation and fate of a molecular excited singlet state, including absorption (ABS), fluorescence (FL), phosphorescence (PH), internal conversion (IC), intersystem crossing (ISC), vibrational relaxation (VR) and collisional quenching (CQ).Not included are processes like solvent relaxation, energy transfer and photochemical reactions
荧光偏振的测量
荧光偏振的测量
Following a pulse excitation, the fluorescence anisotropy of a spherical particle in a homogeneous isotropic medium decays exponentially
发射光谱 激发光谱 时间分辨 空间分辨
内容
荧光及其测量 荧光偏振 应用
– 流动性 – 分子基本性质 – 酶学 – 分子相互作用 – 荧光偏振免疫 – 成像
荧光标记方法
荧光偏振简史
Fluorescence polarization was first described in 1926 by Perrin.
Weber greatly expanded the theory and developed the first instrumentation for the measurement of FP.
Dandliker and co-workers expanded FP into biological systems, such as antigen-antibody reactions and hormone-receptor interactions.