Fluorescence Spectroscopy - the Fluorolog 3.22

Working with polarization

Quantification of the polarization dependency of the secondary monochromator

* Prepare in your cuvette a solution with the chromophore that covers the wavelength of interest.

* Flip the mirror to collect the fluorescence at the angle of 90°.

* Adjust the polarization in excitation to horizontal and the primary monochromator at the right excitation wavelength.

* Record two spectra : one with horizontal polarization and one with vertical polarization in analysis.

* Calculate the ratio of the two spectra. This reflect the different behavior in polarization of the secondary monochromator.

Upper image : Geometrical configuration of the polarizers. Horizontal polarization in excitation and variable in analysis (horizontal and vertical).

Upper image : Emission spectra of NBD-PC in chloroform. In red the polarizer in analysis was set verticaly and in green horizontaly.

Upper image : Intensity ratio of the two spectra.