Improving the fluorescence polarization method to evaluate the orientation of fluorescent systems adsorbed in ordered layered materials

The fluorescence polarization method, recently developed for the evaluation of the preferential orientation of fluorescent dyes adsorbed in layered materials [F. López Arbeloa, V. Martínez Martínez, J. Photochem. Photobiol. A: Chem. 181 (2006) 44], is readapted to improve its application. Fluorescence polarization was previously obtained by recording the emission intensity for two orthogonal orientations of the emission polarizer (i.e., the horizontal and vertical polarized light) after excitation with vertical or horizontal polarized light. In the method proposed in this work, samples are excited with unpolarized light, reducing the polarization effect of the excitation light scattering at those emission wavelengths close to the excitation wavelength. Moreover, the present method decreases the effect of the orientation of other non-fluorescent species present in the system, which are active in the excitation process. Consequently, the new method is more simple, precise and sensitive. It is applied to evaluate the orientation of rhodamine 6G dye adsorbed in ordered laponite clay films with low and moderated dye loadings.