Inner Filter Effect Correction for Fluorescence Measurements in Microplates Using Variable Vertical Axis Focus.

ABSTRACT: The inner filter effect (IFE) hinders fluorescence measurements, limiting linear dependence of fluorescence signals to low sample concentrations. Modern microplate readers allow movement of the optical element in the vertical axis, changing the relative position of the focus and thus the sample geometry. The proposed Z-position IFE correction method requires only two fluorescence measurements at different known vertical axis positions (z-positions) of the optical element for the same sample. Samples of quinine sulfate, both pure and in mixtures with potassium dichromate, showed a linear dependence of corrected fluorescence on fluorophore concentration (R² > 0.999), up to A_ex ≈ 2 and A_em ≈ 0.5. The correction extended linea

SUBMITTER: Weitner T 

PROVIDER: S-EPMC9118198 | biostudies-literature | 2022 May

REPOSITORIES: biostudies-literature