The performance of global (simultaneous) analysis of multiexponential fluorescence decay surfaces using reference convolution is investigated in a systematic way using simulated and experimental data sets. It is shown that the increased model discrimination ability and the more accurate parameter recovery by global analysis as compared to single curve analysis originate from combining decay traces with differing contributions of the decay components. Simultaneous analysis of decay traces in which the contributions of the components are changed as much as possible is the most beneficial. Consequently, including more decay traces in the global decay surface does not necessarily lead to a better model distinction capability. For decay surfaces collected as a function of the emission (excitation) wavelength, the decays with minimal overlap between the emission (absorption) spectra associated with the decay components will contribute the most to the improved model discrimination and parameter recovery.

Date Published: 1 May 1990
PDF: 12 pages
Proc. SPIE 1204, Time-Resolved Laser Spectroscopy in Biochemistry II, (1 May 1990); doi: 10.1117/12.17709

Published in SPIE Proceedings Vol. 1204:
Time-Resolved Laser Spectroscopy in Biochemistry II
Joseph R. Lakowicz, Editor(s)