通过自发闪的光体进行单分子定位扩展纳米镜.
Lu Yang1, Tianli Zhai1, Song Chen1
1Department of Chemistry and Shanghai Key Laboratory of Molecular Catalysis and Innovative Materials Institution, Fudan University, Shanghai 200438, China.
Chemical & biomedical imaging
|February 27, 2026
概括
我们开发了一种新的方法,结合扩展显微镜 (ExM) 和超分辨率显微镜 (STORM),使用一种新型染料. 这种方法克服了以前的局限性,使得细胞结构如线粒体和微管的纳米尺度成像更加清晰.
科学领域:
- 细胞生物学 细胞生物学
- 显微镜技术 显微镜技术
- 生物物理学的生物物理.
背景情况:
- 扩展显微镜 (ExM) 和随机光学重建显微镜 (STORM) 能够对细胞超结构进行超分辨率成像.
- 挑战包括水凝收缩和光体漂白,阻碍了Ex-STORM集成.
研究的目的:
- 通过使用一种新的自发闪染料来克服Ex-STORM的局限性.
- 建立一个精简的工作流程,以提高细胞超结构的纳米尺度成像.
主要方法:
- 使用HMSiR,一种罗达胺染料,在生理pH值下具有内在的随机闪.
- 开发了一个精简的Ex-STORM工作流程,实现稳定的线性扩张.
- 应用该方法可视化线粒体超结构和微管组织.
主要成果:
- 实现了4.2 ± 0.3的稳定线性膨胀系数.
- 已解析的线粒体超结构,包括TOM20蛋白质集群 (~35 nm).
- 可视化微管组织 ~55nm宽度,提供纳米级蛋白质分布见解.
结论:
- HMSiR染料克服了Ex-STORM的局限性,改善了光保留,并消除了专门的缓冲区.
- 精简的Ex-STORM工作流程为细胞超结构成像提供了更高的清晰度.
- 这种方法在细胞生物学中推进了高分辨率成像,使细胞纳米世界的探索成为可能.
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