A FRET pair for quantitative and superresolution imaging of amyloid fibril formation

The presence of neuritic plaques and amyloid fibrils arising from the misfolding of certain proteins is the principal molecular indicator of neurodegenerative diseases such as Alzheimer's and Parkinson's disease. Methodologies for studying the early stages of amyloid aggregation are rapidly arising to provide a better understanding of the mechanism of fibrillization and cytotoxicity and to identify potential targets for diagnosis and therapy. The method presented here involves the simultaneous use of two different fluorophores, a quinolimide derivative and Nile Blue A. These are capable of interacting with and reporting on the formation of preamyloid aggregates and fibrils of apoferritin through fluorescence resonance energy transfer (FRET), which occurs between them, thus maximizing the contrast in detection and quantitative information of such amyloid species by using multidimensional fluorescence lifetime imaging microscopy (FLIM).

Automated summary

The presence of neuritic plaques and amyloid fibrils is key in neurodegenerative diseases, and new methodologies are emerging to study the early stages of amyloid aggregation. This study utilizes two different fluorophores to detect and quantify amyloid species through multidimensional fluorescence lifetime imaging microscopy, providing a better understanding of the mechanism of fibrillization and cytotoxicity.