Phasor-Fluorescence Lifetime Imaging Correlates Dynamics of in-situ Amyloid Nanostructures to Respiratory Dysfunction

The complex dynamics and transience of molecular aggregation pathways leading to amyloid-based neurodegeneration complicate the mechanistic understanding of these fatal diseases. Current technologies are unable to track the molecular processes leading to the onset of amyloidogenesis where real-time information is imperative to correlate its rich biology. Using a chemically-designed amyloidogenic molecule, we map its molecular transformation into amyloids and the resultant fusion with endosomes to form discrete, hollow plaque clusters. Tracked by phasor-fluorescence lifetime imaging (phasor-FLIM) in epithelial cells (L929, A549, MDA-MB 231) and correlative light-electron microscopy/tomography (CLEM), spatiotemporal splicing of the aggregation events shows time-correlated respiratory failure. We reveal that the initial dynamics of aggregation invokes cellular responses on a systemic level.