On the mechanism of Nile blue and Nile red interactions within the ion-selective membrane

Dataset contains fluorescence spectra, absorbance spectra, fluorescence spectra series, FLIM microscopy images and derivative data, from a series of experiments on ion-selective membrane optodes that were acquired by staining a non-conditioned potassium-selective membrane using a water solution of Nile Blue(NB) chloride.The sample that was examined consisted of a regular ion-selective membrane (potassium-selective membrane was used as model). The modified compositions were also used where the membrane would be prepared without either ionophore or ion-exchanger. The membrane would optionally have Nile red (NR) also added to the initial formulation.The experiments that were performed are divided into three parts:First is examination of dye accumulation in the membrane in response to specific membrane compositions (presence/ absence of NR, ionophore, ion-exchanger). It was used to elucidate conditions necessary for the dyes to be incorporated into the membrane. Data contains pictures, absorbance spectra and fluorescence spectra.Second part examines ion-selective membrane of regular composition where all dye present was introduced through exposure to NB chloride water solution. NR and NB are all accumulated in such a membrane, however the fluorescence of NR is quenched/reabsorbed by NB. NB fluorescence is also predominantly excited through these channels rather than directly, by the excitation beam. In response to presence of primary ion the NB is expelled from the membrane which both severs these excitation channels and leads to recovery of NR fluorescence. The effects observed are examined for their applicability in chemical analysis. Data contains fluorescence spectra timeseries (collected over 30 min every 2 minutes) collected for membranes exposed to different concentrations of KCl and also water and interfering ions (Na+). It contains fluorescence spectra of the analyte solutions post measurement and picture om the membrane samples used for color analysis. Derivative data contains fluorescence intensity ratio (600 nm(NR)/ 670 nm(NB)) timetraces, the same ratios are plotted as a function of K+ concentration at different moments.In the third part confocal FLIM microscopy is used to elucidate contributions fluorescence quenching and reabsorption towards suppressing NR fluorescence. The data cotains FLIM images (fluorescence lifetime and intensity intensity) of reference membrane that initially only contained the donor dye, and timeseries (collected over 1h) of FLIM images collected upon the membrane being exposed to nile blue chloride water solution (introduction of the acceptor dye). Data contais tif files, each composed of two images: average fluorescence lifetime and intensity of the donor; over a cross section of the same area of the membrane.