Global methods for time-resolved and steady-state fluorescence and steady-state absorption spectroscopy

Abstract

Global methods for time-resolved and steady-state experimentation and analysis are presented. Examples considered-most of which are nucleic acid systems-include methylated and unmethylated oligo- and polynucleotides formed from alternating C and G bases, for which there is energy transfer; and polynucleotides formed from A and T bases, for which there may also be exciplex formation and dissociation. We show how time-resolved and steady-state data can be used to determine the number and nature of fluorescent components in each system, resolve species-associated absorption and emission spectra (with either time-resolved or steady-state data) , and estimate rates of energy transfer (time-resolved data) or relative quantum yields associated with energy transfer (steady-state data). Where there is an Arrhenius dependence on temperature, energies of activation can also be determined from steady-state data.