FLIM-Based Metabolic Profiling of Endothelial Cells: Differentiating Alzheimer’s Disease from Healthy States

This study examines the metabolic and fluorescence characteristics of endothelial cells from healthy donor and an Alzheimer’s disease patient, providing a novel perspective on cell differentiation under varying photodynamic therapy (PDT) and biochemical conditions. The findings advance the understanding of cellular metabolism's role in disease progression. Fluorescence lifetime imaging microscopy (FLIM) was employed to analyze the autofluorescence of NADH and FAD. Using a femtosecond laser, phasor diagram analysis identified fluorescence lifetime distributions, while photodynamic therapy was applied at varying doses to assess metabolic changes. Distinct fluorescence lifetime parameters for NADH and FAD were observed, with significant differences between donor and Alzheimer’s groups. PDT and biochemical additives affected NADH and FAD signals, demonstrating statistically significant variance using MANOVA. The data also revealed overlapping characteristics under certain conditions, complicating differentiation. This study highlights FLIM’s potential to detect subtle metabolic changes and refine diagnostic approaches in neurodegenerative diseases.