Complex Analysis of Fluorescence Intensity Fluctuations of Molecular Compounds

Abstract

A method is proposed for the complex analysis of fluctuations in the fluorescence intensity of molecular compounds, which allows determining the structural composition of protein oligomers. The idea of the method is to analyze the photon counting histograms of experimental measurements using principal component analysis to assess the presence of oligomeric compounds, and to perform hierarchical cluster analysis, to determine the data classes corresponding to various molecular compounds, followed by selecting cluster medoids to determine the oligomeric composition of protein complexes. The efficiency of the analysis algorithms developed within the framework of the proposed method was confirmed on simulated and experimental photon counting histograms of the measured fluorescence intensity fluctuations of monomeric and dimeric forms of green-fluorescent protein (GFP).