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相关概念视频

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Imaging Biological Samples with Optical Microscopy01:18

Imaging Biological Samples with Optical Microscopy

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Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
In optical microscopy, the specimen to be viewed is placed on a glass slide and clipped on the stage...
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Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

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Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
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Overview of Electron Microscopy01:25

Overview of Electron Microscopy

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The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.
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Visualization of the Immunological Synapse by Dual Color Time-gated Stimulated Emission Depletion STED Nanoscopy
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Visualization of the Immunological Synapse by Dual Color Time-gated Stimulated Emission Depletion STED Nanoscopy

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使用 SUPPOSe 改进 STED 显微镜:从单个图像增强分辨率.

Micaela Toscani1, Axel M Lacapmesure1

  • 1Laboratorio de Fotónica, IIBM-FIUBA, CONICET, Buenos Aires, Argentina.

Methods and applications in fluorescence
|March 8, 2024
PubMed
概括

SUPPOSe算法增强了刺激排放枯竭 (STED) 显微镜,揭示了核孔综合体 (NPC) 的八倍对称性. 这一进步允许从单次显微镜采集中可靠地检索纳米级物体.

科学领域:

  • 细胞生物学 细胞生物学
  • 生物物理学的生物物理.
  • 显微镜的使用方法

背景情况:

  • 核孔综合体 (NPC) 是一个大型蛋白质结构,对核细胞质运输至关重要.
  • NPC特有的环形结构和八重对称性对其功能至关重要.
  • 刺激发射耗尽 (STED) 显微镜可以提供高分辨率成像,但难以解决NPC的完全对称性.

研究的目的:

  • 使用 SUPPOSe 算法来提高 STED 显微镜的分辨率极限.
  • 描述核孔复合体 (NPC) 的纳米结构.
  • 评估 SUPPOSe 算法的可靠性,以检索低分辨率的纳米级物体.

主要方法:

  • 将 SUPPOSe 算法应用于 STED 显微镜对 NPC 中内源标记的 Nup96 的图像.
  • 对562个单个NPC进行分析,以确定结构参数.
  • 概率模型用于评估标签效率和结构兼容性.

主要成果:

  • 假设算法成功地解决了NPC的八边形结构,揭示了它的八倍对称性.
  • 平均NPC半径被确定为R = 54.2 ± 2.9nm.
  • 计算出了31%的有效标签效率,符合单分子定位显微镜标准.
关键词:
子 一个子假设一个假设.基于卷积的算法基于卷积的算法.增强 增强 增强显微镜 显微镜是指使用显微镜.核孔复合体是核孔复合体.超级分辨率的超级分辨率

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结论:

  • 假设算法显著提高了STED显微镜的能力,以解决像NPC这样的复杂纳米结构.
  • 假设可靠地从单个,潜在的杂的显微镜采集中获取分辨率低的纳米级信息.
  • 该方法为生物宏分子的详细结构分析提供了强大的工具.