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相关概念视频

Atomic Force Microscopy01:08

Atomic Force Microscopy

3.6K
Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass, polymers, and biological samples. AFM offers over 1000 times more resolution than the optical imaging system. Images generated from AFM are three-dimensional surface profiles, offering an advantage over the flat, two-dimensional images from other imaging techniques.
The AFM Probe
The probe is regarded as the heart of any AFM setup and comprises the...
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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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Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

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Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

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A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
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相关实验视频

Updated: Sep 10, 2025

Mitochondria and Endoplasmic Reticulum Imaging by Correlative Light and Volume Electron Microscopy
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Mitochondria and Endoplasmic Reticulum Imaging by Correlative Light and Volume Electron Microscopy

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使用相对 AFM/光显微镜成像核包裹

Emilie Costes1, Anthony Vial2, Christine Doucet3

  • 1Centre de Biologie Structurale, INSERM, CNRS, University of Montpellier, Montpellier, France.

Methods in molecular biology (Clifton, N.J.)
|August 20, 2025
PubMed
概括

这项研究提出了一种新方法,用于从哺乳动物细胞中制备核包裹 (NE),保持核孔复合体 (NPC) 结构. 该技术允许使用原子力显微镜 (AFM) 对内部和外部核膜进行高分辨率成像.

关键词:
原子力显微镜 (AFM)直接随机光学重建显微镜 (dSTORM)光显微镜哺乳动物细胞核包裹核孔综合体 (NPC)单分子定位显微镜 (SMLM)

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Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions
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Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions

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Single-Molecule Imaging of Nuclear Transport
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Single-Molecule Imaging of Nuclear Transport

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相关实验视频

Last Updated: Sep 10, 2025

Mitochondria and Endoplasmic Reticulum Imaging by Correlative Light and Volume Electron Microscopy
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Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions
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Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions

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科学领域:

  • 细胞生物学
  • 生物物理
  • 显微镜

背景情况:

  • 核膜 (NE) 是一个具有内外膜的关键真核结构.
  • 由于膜张力损失,现有的NE制备方法可以改变核孔综合体 (NPC) 形态.
  • 原子力显微镜 (AFM) 是用于成像NE结构的强大工具.

研究的目的:

  • 从培养的哺乳动物细胞中制备完整的核包裹 (NE) 的新方法.
  • 在样本制备过程中保持核孔复合物的原生结构.
  • 使用AFM和相关显微镜实现内外核膜的高分辨率成像.

主要方法:

  • 从培养的哺乳动物细胞中提取细胞核并将其贴在玻璃基板上.
  • 温和地打开核以保持膜张力和NPC完整性.
  • 免疫标记结合AFM和直接的随机光学重建显微镜 (dSTORM) 进行相关成像.

主要成果:

  • 这种新的制备方法成功地保持了NE膜张力和NPC结构.
  • 实现内部和外部核膜的高分辨率AFM成像.
  • 相对的AFM/dSTORM成像提供了详细的结构和分子信息.

结论:

  • 该协议为研究具有保存形态的NE和NPC提供了一个强大的方法.
  • 该方法适用于各种细胞模型,包括植物细胞和组织.
  • 它可以提供先进的相关成像技术以获得更深入的生物学见解.