Single-cell analysis of autofluorescence lifetime images

Multiphoton fluorescence lifetime imaging of the metabolic coenzymes reduced nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) allows quantification of cellular metabolism. Due to the link between cellular metabolism and cell function, autofluorescence lifetime imaging provides many features for identification of cells with different phenotypes. Segmentation of multiphoton fluorescence lifetime images allows analysis of data at a single-cell level and quantification of cellular heterogeneity. In this study, Gaussian distribution modeling and machine learning classification algorithms are used for the identification of rare cells within autofluorescence lifetime image data.