Laser-induced fluorescence reader with a turbidimetric system for sandwich-type immunoassay using nanoparticles

Graphical abstract: Laser-induced fluorescence reader with ratiometric correction for sandwich-type immunoassay using nanoparticles. - Highlights: • Laser-induced fluorescence system with ratiometric correction was developed. • The system reduced experimental error caused by particle loss and aggregation. • The detection limit of about 39 pg mL⁻¹ for salinomycin was obtained. • Calibration linearity and sensitivity were also significantly improved. • The system has the potential for bioanalysis using various nanoparticles. - Abstract: A unique laser-induced fluorescence (LIF) reader equipped with a turbidimetric system was developed for a sandwich-type immunoassay using nanoparticles. The system was specifically designed to reduce experimental error caused by particle loss, aggregation and sinking, and to improve analytical performance through ratiometric measurement of the fluorescence with respect to the turbidimetric absorbance. For application to determine the concentration of salinomycin, magnetic nanoparticles (MNPs) and FITC-doped silica nanoparticles (colored balls) immobilized with antibody were synthesized for magnetic extraction and for tagging as a fluorescence probe, respectively. The detection limit of about 39 pg mL⁻¹ was obtained, which was an improvement of about 2-fold compared to that obtained without employment of the turbidimetric system. Calibration linearity and sensitivity were also improved, with increase from 0.8601 to 0.9905 in the R²-coefficient and by 1.92-fold for the curve slope, respectively. The developed LIF reader has the potential to be used for fluorescence measurements using various nanomaterials, such as quantum dots