CORRECTION FOR UNEQUAL INTENSITIES OF LEFT AND RIGHT CIRCULARLY-POLARIZED LIGHT IN STEADY-STATE AND LIFETIME-RESOLVED FLUORESCENCE-DETECTED CIRCULAR-DICHROISM

A major difficulty in fluorescence-detected circular dichroism (FDCD) and lifetime-resolved fluorescence-detected circular dichroism (LRFDCD) is the generation of equal excitation intensities of left circularly polarized light (LCPL) and right circularly polarized light (RCPL). In the presence of unequal intensities, the observed FDCD signal of an optically active sample, or the resolved FDCD signals of a multicomponent system in the case of LRFDCD, will be contaminated by a factor that is the ratio of the two unequal intensities. For optically inactive samples, a sample-independent, artifactual, nonzero signal of constant magnitude is observed. A general scheme is presented for the correction of these inaccuracies caused by unequal intensities of LCPL and RCPL. Large differences between LCPL and RCPL excitation intensities were artificially introduced in steady-state FDCD measurements, and the artifact was accurately corrected by the scheme. Corrected results for the different experimental approaches that have been described for LRFDCD showed similarly good accuracy. In a related consideration, inclusion of the total absorbance and absorption circular dichroism of the sample in the calculation of the FDCD signal is shown to be essential for samples with high absorbances.