TY - GEN
T1 - Long-term, super-resolution imaging of amyloid structures using transient amyloid binding microscopy
AU - Ding, Tianben
AU - Spehar, Kevin
AU - Bieschke, Jan
AU - Lew, Matthew D.
N1 - Funding Information:
Research reported in this publication was supported by the National Science Foundation under grant no. ECCS-1653777 to M.D.L, by the National Institute of General Medical Sciences of the National Institutes of Health under grant no. R35GM124858 to M.D.L., and the Hope Center for Neurological Disorders pilot through a grant to J.B. The authors thank Yuanzi Sun and Niraja Kedia for technical assistance.
Publisher Copyright:
© COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only.
PY - 2019
Y1 - 2019
N2 - 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.
AB - 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.
KW - Amyloid aggregation
KW - Amyloid-beta peptides
KW - Bindingactivated fluorescence
KW - Single-molecule localization microscopy
UR - http://www.scopus.com/inward/record.url?scp=85066755084&partnerID=8YFLogxK
U2 - 10.1117/12.2507656
DO - 10.1117/12.2507656
M3 - Conference contribution
AN - SCOPUS:85066755084
T3 - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
BT - Single Molecule Spectroscopy and Superresolution Imaging XII
A2 - Koberling, Felix
A2 - Gryczynski, Zygmunt K.
A2 - Gregor, Ingo
PB - SPIE
T2 - Single Molecule Spectroscopy and Superresolution Imaging XII 2019
Y2 - 2 February 2019 through 3 February 2019
ER -