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自发闪的探测器用于在活细胞中无洗的基于单分子定位的超分辨率成像
Qinglong Qiao1, Aoxuan Song1,2, Kai An1,2
1Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian, 116023, China.
Angewandte Chemie (International ed. in English)
|November 13, 2024
概括
一个新的blinkogenic探头,HM-DS655-Halo,可以在活细胞中进行超分辨率成像. 它保持关闭,直到与HaloTag结合,显著减少背景噪声,以精确可视化细胞结构.
科学领域:
- 生物物理学的生物物理.
- 细胞生物学 细胞生物学
- 显微镜的使用方法
背景情况:
- 单分子定位超分辨率光成像 (SMLM) 依赖于探针闪.
- 没有反应的探头会引起背景噪声,降低成像准确度.
- 现有的探头需要冲洗的步骤,以获得有效的SMLM.
研究的目的:
- 开发一种新的blinkogenic探针,用于在活细胞中直接检测SMLM.
- 为了使SMLM没有探头冲洗的步骤.
- 为了提高SMLM中的定位精度和准确性.
主要方法:
- 开发了HM-DS655-Halo,这是一个通过HaloTag结合激活的闪性探针.
- 引入了RDC参数来量化闪性.
- 在活细胞的直接SMLM成像中应用了探头.
主要成果:
- HM-DS655-Halo从OFF过渡到闪HaloTag绑定时的情况.
- 12的RDC值表示超级闪性.
- 在活细胞SMLM中实现了有效的背景抑制.
- 对线粒体,细胞迁移和伪足的动态成像是成功的.
结论:
- HM-DS655-Halo促进了活细胞中直接的SMLM,而无需洗.
- 探测器的自动闪活动显著降低了背景噪音.
- 这一进步提高了动态细胞成像的精度和准确性.
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