Reducing the Inner Filter Effect in Microplates by Increasing Absorbance? Linear Fluorescence in Highly Concentrated Fluorophore Solutions in the Presence of an Added Absorber.

ABSTRACT: The fluorescence attenuation caused by the absorption of the excitation and/or emission light is called the Inner Filter Effect (IFE) and can lead to a nonlinear fluorescence concentration response. In this article, we propose the AddAbs (Added Absorber) method, which counterintuitively corrects IFE by increasing the absorbance of the sample. In this method, an equal amount of a highly absorbing chromophore is added to each sample to compensate for the nonuniform quenching caused by different fluorophore concentrations. The AddAbs method was able to provide a linear fluorescence response (R² > 0.999) for very concentrated fluorophore solutions with extreme IFE over more than 97% of the concentration range with less than 1% deviation in calibration slope. The true limit fo

SUBMITTER: Friganovic T 

PROVIDER: S-EPMC10483458 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature