Method for measuring response function of time resolution fluorescence instrument in yellow-green optical band

A technology of instrument response function and measurement time, applied in fluorescence/phosphorescence, material excitation analysis, etc., can solve problems such as instrument response function system error, and achieve the effects of improving accuracy, large quantum yield, and easy preparation

Inactive Publication Date: 2015-02-11
EAST CHINA NORMAL UNIV
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Problems solved by technology

When using this method to obtain the instrument response function, there are some defects. For example, the signal can only be detected when the detection wavelength is close to the excitation wavelength. However, the wavelength of the fluorescence emitted by the excited substance has a certain deviation from the wavelength of the excitation light. shift, and there are color effects in measuring instruments such as monochromators, photomultiplier tubes or avalanche photodiodes, which will lead to systematic errors in the instrument response function obtained by this method

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  • Method for measuring response function of time resolution fluorescence instrument in yellow-green optical band
  • Method for measuring response function of time resolution fluorescence instrument in yellow-green optical band
  • Method for measuring response function of time resolution fluorescence instrument in yellow-green optical band

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[0029] The specific implementation manner of the present invention will be further described in detail below in conjunction with the accompanying drawings and examples, but the protection scope of the present invention should not be limited thereby.

[0030] Preparation of reference dyes

[0031] First prepare the disodium hydrogen phosphate-sodium hydroxide buffer solution with pH 10, weigh 18.3g of sodium iodide (NaI), dilute it to 10mL with the prepared pH 10 buffer solution, and prepare 12.2M (nearly saturated) NaI Solution; weigh 1.91g of sodium fluorescein, and dilute to 100mL with the prepared pH10 buffer solution to prepare a 51.4mM sodium fluorescein alkaline solution; measure the prepared 12.2M NaI (sodium iodide) solution 1000 μL, 10 μL of 51.44 mM sodium fluorescein solution and mix well, the obtained mixed solution is the reference dye for measuring the instrument response function of time-resolved fluorescence, wherein the final concentration of iodide ion as a...

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Abstract

The invention aims to provide a method for measuring a response function of a time resolution fluorescence instrument in the yellow-green optical band. The method comprises the following steps: a, using quenched fluorescein sodium as a reference dye, using fluorescein sodium as a fluorescent dye, and using iodine ion (I<->) as a quenching agent; b, acquiring the time resolution fluorescence spectrum of the reference dye through a measurement instrument; and c, using the time resolution fluorescence spectrum as the response function of the measurement instrument. The final concentration of fluorescein sodium is 200-800 muM, the final concentration of iodide ion (I<->) is 8-12.2 M, and the pH value of the system is more than or equal to 9. Compared with the method of solid particle scattering measurement instrument response function, the method of the invention to obtain the instrument response function avoids color effect between the different excitation wavelengths, improves the result accuracy of the whole experiment, and avoids the trouble of replacing filter.

Description

technical field [0001] The invention belongs to the field of optical spectrum measurement, and relates to a method for measuring the response function of a time-resolved fluorescence instrument, in particular to a method for measuring the response function of a time-resolved fluorescence instrument in the yellow-green light band. Background technique [0002] In time-resolved fluorescence spectroscopy, the measured fluorescence decay curve is actually the convolution of the instrument response function (Instrument Response Function, IRF) and the ideal time decay curve. To obtain an ideal time decay curve, it is necessary to remove the influence of the instrument response function (IRF). Therefore, whether the instrument response function is accurate will directly affect the measurement accuracy of the ideal time decay curve. Typically, the instrument response function is obtained by measuring the time-domain profile of the excitation light. Rayleigh light scattering provid...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 潘海峰贾梦辉刘梦薇李磊赵力涛常孟方张三军徐建华
Owner EAST CHINA NORMAL UNIV
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