自动闪的蒂奥夫拉T用于超分辨率成像
Qiqi Yang1, Elnaz Hosseini1, Peigen Yao1
1Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany.
The journal of physical chemistry letters
|July 19, 2024
概括
提奥夫拉T (ThT) 染料对单分子超分辨率成像表现出固有的随机闪,这与之前的看法相反. 这种特性使得无需专门的缓冲器,可以可视化粉样纤维.
科学领域:
- 生物物理学的生物物理.
- 化学生物学是化学生物学.
- 显微镜的使用方法
背景情况:
- 提奥夫拉T (ThT) 是一种用于检测粉样和类纳米纤维的标准染料.
- 一般认为ThT在结合聚合蛋白时具有稳定的光.
- 单分子定位显微镜 (SMLM) 需要光交换/闪特性来进行超分辨率成像.
研究的目的:
- 在单个分子水平上研究ThT的内在光交换/闪特性.
- 为了确定ThT是否可以在没有专门的闪缓冲器的情况下用于SMLM.
- 在各种实验条件下描述ThT的闪行为.
主要方法:
- 单分子光谱学分析ThT光.
- 在超低度下研究ThT闪特性 (光子数量,持续时间,开关比).
- 在空气和PBS缓冲器中对THT标记的α-synuclein纤维的SMLM成像.
主要成果:
- 在稳定的约束条件下,ThT显示了内在的随机闪,这与之前的假设相矛盾.
- 在单个分子水平上量化了ThT的闪特征.
- 使用ThT.成功地实现了α-synuclein纤维的SMLM成像.
结论:
- ThT具有固有的光交换能力,适合SMLM.
- 基于ThT的粉样结构的SMLM不需要专门的闪缓冲器.
- 这一发现扩大了ThT在蛋白质聚合物的超分辨率显微镜中的实用性.
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