Characterization of early-stage amyloid aggregates by incorporating extrinsic fluorescence and atomic force microscopy.

Amyloid-β (Aβ) oligomers are nanosized bio-assemblies that cause Alzheimer's disease. Characterizing early-stage Aβ aggregates becomes an important issue because it is a prerequisite in exploring small molecule inhibitors that bind to Aβ oligomers. Of special interest are efficient screening systems that characterize the Aβ oligomer size with respect to the aging time. In this work, highly sensitive fluorescence techniques and atomic force microscopy (AFM) were employed to investigate the size determination of Aβ and screening of small molecule inhibitors. A solvatochromic dye, 1-anilinonaphthalene-8-sulfonic acid (ANS), was used as an extrinsic fluorophore to monitor the growth mechanism of the Aβ aggregates. Then, the time-resolved fluorescence anisotropy method was employed to estimate the hydrodynamic size of Aβ oligomers. Finally, AFM was used to characterize the Aβ oligomer size in the absence and presence of potential inhibitors. We present that the combination of such three experimental techniques is an excellent way to detect the early stage of Aβ aggregation and to screen small molecule inhibitors.