The Concept of λ-Ratiometry in Fluorescence Sensing and Imaging

被引:170
作者
Demchenko, Alexander P. [1 ]
机构
[1] Natl Acad Sci Ukraine, Palladin Inst Biochem, UA-01030 Kiev, Ukraine
关键词
Intensity sensing; Anisotropy; Time-resolved fluorimetry; Wavelength ratiometry; Excited-state reactions; INTRAMOLECULAR PROTON-TRANSFER; RESONANCE ENERGY-TRANSFER; EXCITED-STATE REACTIONS; EMISSION PH SENSORS; CHARGE-TRANSFER; DUAL-EMISSION; EXCIMER FLUORESCENCE; CONJUGATED POLYMERS; ELECTRON-TRANSFER; NUCLEIC-ACIDS;
D O I
10.1007/s10895-010-0644-y
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Very limited number of parameters is available for fluorescence sensing and imaging. The changes of intensity are of low analytical value due to the absence of internal reference. Anisotropy and lifetime sensing have their own limitations. In this respect the lambda-ratiometric (based on intensity ratios at two or more wavelengths) recording of spectral changes becomes more popular. Because the spectral changes are connected directly with the variations of interaction energies this approach is seen as the most universal method to study intermolecular interactions. It is applicable for different sensor formats and for obtaining analytical information from cell images. Here we critically analyze different approaches in lambda-ratiometric sensing that use single and double fluorescence emitters and are based on different mechanisms producing spectroscopic change. Very promising is the exploration of mechanisms that allow obtaining ratiometric response from a single dye.
引用
收藏
页码:1099 / 1128
页数:30
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