Department of Biomedical Engineering, Washington University in St. Louis, St. Louis, MO, 63130, USA.
Department of Electrical and Systems Engineering, Washington University in St. Louis, St. Louis, MO, 63130, USA.
Chembiochem. 2018 Sep 17;19(18):1944-1948. doi: 10.1002/cbic.201800352. Epub 2018 Aug 8.
Oligomeric amyloid structures are crucial therapeutic targets in Alzheimer's and other amyloid diseases. However, these oligomers are too small to be resolved by standard light microscopy. We have developed a simple and versatile tool to image amyloid structures by using thioflavin T without the need for covalent labeling or immunostaining. The dynamic binding of single dye molecules generates photon bursts that are used for fluorophore localization on a nanometer scale. Thus, photobleaching cannot degrade image quality, allowing for extended observation times. Super-resolution transient amyloid binding microscopy promises to directly image native amyloid by using standard probes and record amyloid dynamics over minutes to days. We imaged amyloid fibrils from multiple polypeptides, oligomeric, and fibrillar structures formed during different stages of amyloid-β aggregation, as well as the structural remodeling of amyloid-β fibrils by the compound epi-gallocatechin gallate.
寡聚体淀粉样结构是阿尔茨海默病和其他淀粉样疾病的重要治疗靶点。然而，这些寡聚体太小，无法用标准的光学显微镜来解析。我们开发了一种简单而通用的工具，通过使用硫黄素 T 来成像淀粉样结构，而不需要共价标记或免疫染色。单个染料分子的动态结合会产生光脉冲，用于在纳米尺度上定位荧光团。因此，荧光漂白不会降低图像质量，从而可以进行更长时间的观察。超分辨率瞬态淀粉样结合显微镜有望使用标准探针直接对天然淀粉样蛋白成像，并记录淀粉样蛋白在数分钟到数天内的动态变化。我们对来自多种多肽的淀粉样纤维、寡聚体和在淀粉样蛋白-β聚集的不同阶段形成的纤维状结构，以及化合物表没食子儿茶素没食子酸酯对淀粉样蛋白-β纤维的结构重塑进行了成像。
