Fluorescence detection by intensity change based sensors: a theoretical model

According to Fluorescence Detection by Intensity Changes (FDIC) the fluorescence intensity of many fluorophores depends on the non-covalent (specific and/or non-specific) interactions these fluorophores would be able to establish with the solvent and, more interestingly, with other surrounding molecules. This latter effect is the basis of FDIC for analytical purposes. In this paper, a preliminary study of FDIC applications using a fluorophore supported in a solid medium (sensor film) is presented. First, a mathematical model relating the analyte concentration with the immobilized fluorophore fluorescence is deduced. The model includes all the different mechanisms explaining this relationship: index of refraction or dielectric constant modification, scattering coefficient alteration and sensor film volume increase. Then, the very first experimental results are presented, using different fluorophores and solid supports. The best results were obtained using polyacrylamide (PAA) polymers and coralyne as the fluorophore. This sensor film is applied for albumin and polyethylenglycol determination and the results are compared with those obtained using coralyne in solution. Albumin quenches the coralyne fluorescence in both cases (solution and film), while PEG quenches coralyne fluorescence in films but increases it in solution. These results suggest that the outstanding fluorescence change mechanism is sensor films is the film volume increases, which is different than those observed in solution.