扩展和波动增强显微镜用于细胞和组织的纳米级分子分析
Dominik Kylies1,2, Hannah S Heil3,4, Arturo G Vesga3,5
1III. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Nature protocols
|July 3, 2025
概括
这项研究介绍了一种可访问的方法,它结合了扩展显微镜和增强的超分辨率辐射波动 (eSRF),以实现传统显微镜的纳米级成像分辨率,使先进的细胞可视化得到广泛应用.
科学领域:
- 细胞生物学 细胞生物学
- 显微镜的使用方法
- 生物医学研究生物医学研究
背景情况:
- 超分辨率显微镜可以提供纳米级分子分析,但需要专门的,昂贵的设备.
- 传统的光显微镜受到衍射极限的限制,限制了详细的细胞可视化.
研究的目的:
- 提供一个详细的协议,以实现纳米级分辨率,使用可访问的显微镜技术.
- 将扩展显微镜与增强的超分辨率辐射波动 (eSRF) 结合起来,以改善成像.
主要方法:
- 使用膨胀显微镜物理放大样本.
- 通过ESRF分析光波动来计算增强图像分辨率.
- 详细说明样品准备,显微镜优化和质量控制.
主要成果:
- 使用常规衍射有限显微镜实现纳米分辨率 (25nm).
- 展示了一种灵活和可访问的方法,用于用多个标签对各种样本类型进行成像.
- 提供了定量工具 (NanoJ-SQUIRREL) 来评估解决方案和忠诚度.
结论:
- 扩展增强的eSRF显微镜为昂贵的超高分辨率技术提供了切实可行的替代方案.
- 能够详细研究细胞超结构,蛋白质定位和相互作用.
- 适用于细胞生物学,组织病理学和更广泛的生物医学研究应用.
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