Fluorescence lifetime distributions in membrane systems

Membranes are complex biological systems that display heterogeneity at all spatial scales. At a molecular level, the heterogeneity arises from lipid and protein composition. At the cellular level, heterogeneity is due to membrane organization and large scale morphology. A quantitative evaluation of membrane heterogeneity at a microscopic level is very important for several fields of membrane studies. We have developed a method for the analysis of the decay of fluorescent membrane probes that can provide a quantity sensitive to membrane heterogeneity. This method is based on the analysis of the fluorescence decay using continuous lifetime distributions. The major challenge in the interpretation of the analysis results is in the identification, at a molecular level, of the mechanisms that influence the fluorescence decay. In this review we illustrate the principles of data analysis and we show examples of identification of the measured parameters with specific variables that affect membrane heterogeneity.