Studies of the growth, evolution, and self-aggregation of β-amyloid fibrils using tapping-mode atomic force microscopy.

Amyloid peptide (Aβ) is the major protein component of plaques found in Alzheimer's disease, and the aggregation of Aβ into oligomeric and fibrillic assemblies has been shown to be an early event of the disease pathway. Visualization of the progressive evolution of nanoscale changes in the morphology of Aβ oligomeric assemblies and amyloid fibrils has been accomplished ex situ using atomic force microscopy (AFM) in ambient conditions. In this report, the size and the shape of amyloid β(1-40) fibrils, as well as the secondary organization into aggregate structures were monitored at different intervals over a period of 5 months. Characterizations with tapping-mode AFM serve to minimize the strong adhesive forces between the probe and the sample to prevent damage or displacement of fragile fibrils. The early stages of Aβ growth showed a predominance of spherical seed structures, oligomeric assemblies, and protofibrils; however the size and density of fibrils progressively increased with time. Within a few days of incubation, linear assemblies and fibrils became apparent. Over extended time scales of up to 5 months, the fibrils formed dense ensembles spanning lengths of several microns, which exhibit interesting changes due to self-organization of the fibrils into bundles or tangles. Detailed characterization of the Aβ assembly process at the nanoscale will help elucidate the role of Aβ in the pathology of Alzheimer's disease.