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相关概念视频

Cryo-electron Microscopy01:28

Cryo-electron Microscopy

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Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...
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Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

2.4K
Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
Electron Tomography
Electron tomography can be performed either in TEM or STEM (scanning transmission...
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Updated: Jun 28, 2025

Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction
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Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction

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使用冷电子显微镜确定宏分子结构

Pradeep Hiriyur Nagaraj1

  • 1Institute of Molecular Biotechnology, Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU), Vienna, Austria. pradeep.hiriyur.nagaraj@bath.edu.

Methods in molecular biology (Clifton, N.J.)
|April 24, 2024
PubMed
概括

低温电子显微镜 (cryo-EM) 提供了对植物蛋白和复合体的高分辨率结构洞察力. 本指南为初学者介绍了冷电磁和负染色电子显微镜 (NSEM) 的样本准备和数据分析.

科学领域:

  • 结构生物学是结构生物学.
  • 生物物理学的生物物理.
  • 植物分子生物学 植物分子生物学

背景情况:

  • 对宏分子和蛋白质复合体的高分辨率结构洞察力对于理解分子功能至关重要.
  • 低温电子显微镜 (cryo-EM) 是一种强大的技术,用于研究原生或近原生状态的结构.
  • 植物生物学家对采用冷EM进行生物分子相互作用研究表示犹,导致数据库中植物衍生结构较少.

研究的目的:

  • 引入用于植物蛋白和宏分子复合物的负染色电子显微镜 (NSEM) 和冷EM的样品制备技术.
  • 为植物结构生物学初学者提供使用单颗粒分析 (SPA) 的数据分析指南.
  • 鼓励在植物科学中应用冷EM,以更深入地了解分子机制.

主要方法:

  • 负染色电子显微镜 (NSEM) 用于样品制备.
  • 低温电子显微镜 (cryo-EM) 用于高分辨率的结构确定.
  • 单粒子分析 (SPA) 用于数据处理和结构重建.

主要成果:

  • 化EM显著提高了对光合作用,能量转移和植物病毒的理解.
  • 尽管采用速度缓慢,但冷EM提供了对植物蛋白功能的详细结构洞察.
  • 该章节详细介绍了样本准备和数据分析的实际步骤,使冷EM更容易获得.
关键词:
低温电子显微镜的低温电子显微镜.图像处理 图像处理负染色电子显微镜的电子显微镜.蛋白质结构 蛋白质结构样品的准备 样品的准备一个粒子分析分析.三维 (3D) 重建的重建.

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Determination of Molecular Structures of HIV Envelope Glycoproteins using Cryo-Electron Tomography and Automated Sub-tomogram Averaging

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Last Updated: Jun 28, 2025

Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction
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Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction

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Single Particle Cryo-Electron Microscopy: From Sample to Structure
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Single Particle Cryo-Electron Microscopy: From Sample to Structure

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Determination of Molecular Structures of HIV Envelope Glycoproteins using Cryo-Electron Tomography and Automated Sub-tomogram Averaging
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结论:

  • 低温电磁波是阐明植物生物过程的分子基础的重要工具.
  • 可访问的样本准备和数据分析协议可以克服植物科学中冷EM采用的障碍.
  • 增加冷EM的应用将加速植物分子生物学和功能方面的发现.