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相关概念视频

Electron Microscope Tomography and Single-particle Reconstruction01:07

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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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The endoplasmic reticulum or ER makes up for more than half of the membranes in a cell and accounts for 10% of total cell volume. It is also the primary protein and lipid synthesis factory for most cell organelles, such as the Golgi apparatus, lysosomes, secretory vesicles, and the plasma membrane. Despite being the most extensive and functionally complex subcellular organelle, ER was the last to be discovered. After years of deliberation, Keith Porter and George Palade in the year 1954,...
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Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
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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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相关实验视频

Updated: Jul 2, 2025

Mitochondria and Endoplasmic Reticulum Imaging by Correlative Light and Volume Electron Microscopy
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电子显微镜技术用于3D植物ER成像3D成像

Charlotte Pain1, Maike Kittelmann2

  • 1Endomembrane Structure and Function Research Group, Biological and Medical Sciences, Oxford Brookes University, Oxford, UK.

Methods in molecular biology (Clifton, N.J.)
|February 27, 2024
PubMed
概括

本研究介绍了两种3D电子显微镜协议,用于可视化植物内质网膜 (ER). 这些方法允许使用ZIO染色或高压结技术详细分析根髓系统细胞中的ER结构.

关键词:
3D EM 3D EM 3D 3D EM 3D 3D 3D 3D 3D 3D 3D 3D 3D 3D 3D 3D 3D 3D 3D 3D自动化细分系统 自动化细分系统细胞内膜网膜的内细胞网膜.高压结方式的高压结方式.序列块面面是SEM的,可以用SEM.齐奥 齐奥 齐奥 齐奥 齐奥

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科学领域:

  • 植物细胞生物学 植物细胞生物学
  • 显微镜技术 显微镜技术

背景情况:

  • 细胞内膜网膜 (ER) 形成了一个广泛的细胞网络,对植物细胞功能至关重要.
  • 了解ER的空间组织,特别是根介质系统,需要高分辨率的成像.
  • 现有的方法缺乏足够的分辨率,无法在植物组织中进行详细的ER结构分析.

研究的目的:

  • 开发和介绍两种用于3D电子显微镜 (EM) 绘制植物ER的新型协议.
  • 为了能够在植物细胞中的各种尺度上详细分析ER结构.
  • 为了促进ER结构的自动或半自动细分.

主要方法:

  • 协议1:选择性ER染色与ZIO进行大规模ER结构分析和自动细分.
  • 协议2:高压冷用于植物组织的快速固定,适用于对较小的ER区域进行详细分析,无需特殊染色.
  • 将技术应用于各种成像模式,包括SBF-SEM和FIB-SEM.

主要成果:

  • 通过使用开发的协议,证明了植物细胞中ER网络的成功可视化.
  • 在根髓系统细胞中实现了ER结构的高分辨率3D成像.
  • 验证了ZIO染色和高压冷对各种EM应用的适用性.

结论:

  • 提出的3D EM协议显著推进了对工厂ER空间组织的研究.
  • 这些技术为植物细胞中ER的详细结构分析提供了强大的工具.
  • 这些方法可以适应各种成像模式,在植物科学研究中具有广泛的应用.