Two-photon excitation of emissive markers with near-infrared (NIR) light is of a particular interest for imaging in biology and medicine because NIR light is relatively weakly absorbed and scattered by tissues. At the same time the mechanism of two-photon absorption allows excitation of molecules located deep inside a scattering medium. In this work we demonstrate that the two-photon excitation combined with the effect of light amplification in the stimulated emission process provides a sensitive method for detecting amyloids of different forms. We investigate the two-photon excited amplified spontaneous emission (ASE) of a fluorescent dye, coumarin 307, in the brain tissue infiltrated with various amyloid phantoms i.e. oligomers, protofibrils and mature fibrils. All these forms of amyloids can be detected by observation of ASE and determination of thresholds for light amplification. On this basis we suggest that a relatively simple extension of currently used emission-based optical spectroscopy techniques can provide key information on pathogenic amyloid structures in tissue.
Keywords: Amplified spontaneous emission; Coumarin; Neurodegeneration; Protein aggregation; Stimulated emission; Time-resolved fluorescence; Two-photon absorption.
Copyright © 2022 Elsevier B.V. All rights reserved.