Using Fractional Intensities of Time-resolved Fluorescence to Sensitively Quantify NADH/NAD⁺ with Genetically Encoded Fluorescent Biosensors.

ABSTRACT: In this paper, we propose a novel and sensitive ratiometric analysis method that uses the fractional intensities of time-resolved fluorescence of genetically encoded fluorescent NADH/NAD⁺ biosensors, Peredox, SoNar, and Frex. When the conformations of the biosensors change upon NADH/NAD⁺ binding, the fractional intensities (α _i τ _i ) have opposite changing trends. Their ratios could be exploited to quantify NADH/NAD⁺ levels with a larger dynamic range and higher resolution versus commonly used fluorescence intensity and lifetime methods. Moreover, only one excitation and one emission wavelength are required for this ratiometric measurement. This eliminates problems of traditional excitation-ratiometric and emission-ratiometric methods.

SUBMITTER: Chang M 

PROVIDER: S-EPMC5482812 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature