Global Analysis of Time-Resolved Fluorescence Data

In this chapter, we describe the global analysis approach for processing time-resolved fluorescence spectroscopy data of molecules in the condensed phase. Combining simultaneous analysis of data measured under different experimental conditions (spatial coordinates, temperature, concentration, emission wavelength, excitation intensity, etc.) with the fitting strategy, enabling parameter linkage and thus decreasing the total amount of estimated variables, makes global analysis more robust and more consistent compared to a sequential fit of single experimental data. We consider the main stages of the global analysis approach and provide some details that are important for its practical implementation. The application of the global approach to the analysis of time-resolved fluorescence anisotropy is demonstrated on experimental data of (enhanced) green fluorescent protein in aqueous solution.