Quantitative measurement method for fluorescence lifetime and fluorescence dynamic anisotropic parameters

An anisotropy and fluorescence lifetime technology, applied in the field of fluorescence characteristics measurement of fluorescent samples, can solve the problem that no measurement method is given, and the measurement method cannot effectively quantitatively measure the fluorescence lifetime.

Inactive Publication Date: 2014-04-02
GUANGDONG UNIV OF TECH
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[0007] Jameson.D.M, Gartton, E.and Hall.R, "The measurement and analysis of heterogeneous emissions by multifrequency phase and modulation fluorometry", Appl. Spectrosc. Rev. 20.55-106 (1984) and Joseph R. Lakowicz, Principles of Fluorescence Spectroscopy.(Springer, New York, 2006), 3rd ed proposed a method for quantitatively measuring the fl

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  • Quantitative measurement method for fluorescence lifetime and fluorescence dynamic anisotropic parameters
  • Quantitative measurement method for fluorescence lifetime and fluorescence dynamic anisotropic parameters
  • Quantitative measurement method for fluorescence lifetime and fluorescence dynamic anisotropic parameters

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[0039] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

[0040] Rhodamine 6G is a commonly used material in bioluminescent labeling technology and is widely used in DNA and protein research. The existing sample solution is a mixture of 10 μM rhodamine 6G and 91% aqueous glycerin solution, and its fluorescence lifetime and fluorescence anisotropy measurement devices are as follows: figure 1 As shown, in this example, the sample solution is excited with a 100 fs pulsed polarized excitation light from the x direction parallel to the z axis, with I ∏ (t) represents the time-resolved polarized fluorescence intensity parallel to the pulsed polarized excitation light, represented by I ⊥ (t) represents the time-resolved polarized fluorescence intensity perpendicular to the pulsed polarized excitation light. The total fluorescence intensity I ∑ (t) is,

[0041] I ∑ (t)=I ∏ (t)+2I ⊥ (t)=I 0 -ex...

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Abstract

The invention relates to a measuring method for fluorescence characteristics of a fluorescence product and particularly relates to a method for quantitative measurement method for fluorescence lifetime and fluorescence dynamic anisotropic parameters. The method comprises the steps of: exciting a product by a pulse or sine polarization exciting light to produce fluorescence; detecting intensity of polarized fluorescence which is parallel to the pulse or sine polarization exciting light and intensity of polarized fluorescence which is perpendicular to the pulse or sine polarization exciting light respectively with a photoelectric detector; and calculating the average life, the fluorescence anisotropy at the initial time, the fluorescence anisotropy at the infinite time and fluorescence molecule rotational correlation time of the sample. With the adoption of the method provided by the invention, the fluorescence lifetime of all samples can be effectively and quantitatively measured; moreover, the invention provides a quantitative measurement method for fluorescence parameters of the fluorescence product, so the method has great application value for quantitative study of the interaction between large biological molecules, such as proteins, in the living cell.

Description

technical field [0001] The invention relates to a method for measuring fluorescence characteristics of fluorescent samples, in particular to a method for quantitatively measuring fluorescence lifetime and fluorescence dynamic anisotropy parameters. Background technique [0002] Due to the emergence of fluorescent protein molecules (FP), the use of fluorescent labeling and fluorescence measurement techniques to study the biophysical chemistry of living cells and biological tissues has become one of the hotspots in life science research. With the development of modern optics and computer technology, it has been discovered that fluorescence anisotropy and fluorescence lifetime measurement techniques can break through the limitation of Abbe's principle and characterize the interaction between biological macromolecules inside living cells. As described by F.S.Wouters, "The physics and biology of fluorescence microscopy in the life science", in Contemporary Physics (2006), Vol.47,...

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Application Information

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IPC IPC(8): G01N21/64
Inventor 周延周白玉磊王钦若
Owner GUANGDONG UNIV OF TECH
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