Long-term, super-resolution imaging of amyloid structures using transient amyloid binding microscopy
Amyloid fibrils and tangles are signatures of Alzheimer disease, but nanometer-sized aggregation intermediates are hypothesized to be the structures most toxic to neurons. The structures of these oligomers are too small to be resolved by conventional light microscopy. We have developed a simple and versatile method, called transient amyloid binding (TAB), to image amyloid structures with nanoscale resolution using amyloidophilic dyes, such as Thioflavin T, without the need for covalent labeling or immunostaining of the amyloid protein. Transient binding of ThT molecules to amyloid structures over time generates photon bursts that are used to localize single fluorophores with nanometer precision. Continuous replenishment of fluorophores from the surrounding solution minimizes photobleaching, allowing us to visualize a single amyloid structure for hours to days. We show that TAB microscopy can image both the oligomeric and fibrillar stages of amyloid-β aggregation. We also demonstrate that TAB microscopy can image the structural remodeling of amyloid fibrils by epi-gallocatechin gallate. Finally, we utilize TAB imaging to observe the non-linear growth of amyloid fibrils.
Ding, Tianben; Spehar, Kevin; Bieschke, Jan; and Lew, Matthew D., "Long-term, super-resolution imaging of amyloid structures using transient amyloid binding microscopy" (2019). Electrical & Systems Engineering Publications and Presentations. 6.
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Tianben Ding, Kevin Spehar, Jan Bieschke, Matthew D. Lew, "Long-term, super-resolution imaging of amyloid structures using transient amyloid binding microscopy," Proc. SPIE 10884, Single Molecule Spectroscopy and Superresolution Imaging XII, 108840J (22 February 2019); doi: 10.1117/12.2507656