Use of a stable and solvent-soluble variant of Amylo-Glo, Amylo-Glo+, for the high-contrast and resolution fluorescent localization of amyloid plaques in murine and human brain tissue sections

Abstract

BackgroundAlthough numerous different fluorescent compounds have been used for the fluorescent localization of amyloid plaques, limitations include weak fluorescence, low contrast, low resolution, broad excitation / emission profile and chemical instability. New methodAmylo-Glo+ was reconstituted from a powder into a stable solution for the histological staining of plaques in brain tissue sections from both patients diagnosed with Alzheimer’s disease and APP/PS1 transgenic mice. ResultsAmylo-Glo+ was found to result in high resolution and contrast labeling of amyloid plaques. The tracer was excited exclusively by ultraviolet light illumination. The staining of amyloid plaques corresponded with the labeling pattern seen following staining with HQ-O. Counterstaining with ethidium bromide resulted in no bleed-through of the Amylo-Glo+ under blue or green light excitation. The tracer was stable in both powder and solution form. Comparison with existing methodsAmylo-Glo+ was found to be more stable, in both a powder and solution form, than the original Amylo-Glo, brighter than Congo red, of narrower emission and greater contrast than Thioflavine S and of narrower emission than X-34 and BSB. ConclusionAmylo-Glo+ is a stable and solvent soluble variant of Amylo-Glo that allows for the high contrast and resolution localization of amyloid plaques in tissue sections from the brains of transgenic mice and humans with Alzheimer’s disease. The tracer is also solely excited by ultraviolet light, making it ideal for fluorescent multiple labeling studies in combination with commonly used red or green fluorophores. It is also stable in both solution and powder form, unlike the original Amylo-Glo which is only available as a basic solution that is prone to precipitation.