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相关概念视频

X-ray Diffraction of Biological Samples01:10

X-ray Diffraction of Biological Samples

3.8K
X-ray diffraction or XRD is an analytical tool that utilizes X-rays to study ordered structures such as crystalline organic and inorganic samples, polycrystalline materials, proteins, carbohydrates, and drugs.
According to Bragg's law, when X-rays strike the sample positioned on a stage, the rays are  scattered by the electron clouds around the sample atoms. The  X-ray diffraction or scattering is caused by constructive interference of the X-ray waves that reflect off the internal...
3.8K
Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

12.3K
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...
12.3K
Cryo-electron Microscopy01:28

Cryo-electron Microscopy

3.2K
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...
3.2K
Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

2.0K
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...
2.0K
Determination of Crystal Structures01:29

Determination of Crystal Structures

135
In the late 1800s, the revelation that light extended beyond visible wavelengths led to the discovery of X-rays by Wilhelm Roentgen. Recognized as high-energy electromagnetic radiation with short wavelengths, X-rays prompted exploration into their interaction with crystals. Max von Laue proposed in 1912 that the periodic arrangement of atoms, ions, or molecules in crystals would cause them to diffract X-rays, a hypothesis confirmed through experiments with copper sulfate and zinc sulfide...
135

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相关实验视频

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Assessing Two-dimensional Crystallization Trials of Small Membrane Proteins for Structural Biology Studies by Electron Crystallography
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Assessing Two-dimensional Crystallization Trials of Small Membrane Proteins for Structural Biology Studies by Electron Crystallography

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超分辨率生物分子晶体学与低分辨率数据.

Gunnar F Schröder1, Michael Levitt, Axel T Brunger

  • 1Institut für Strukturbiologie und Biophysik (ISB-3), Forschungszentrum Jülich, 52425 Jülich, Germany. gu.schroeder@fz-juelich.de

Nature
|April 9, 2010
PubMed
概括

这项研究引入了一种新的方法,用于确定大型生物组件的原子结构,使用低分辨率的X射线衍射数据. 该技术利用已知的同类结构来提高模型的准确性,从而从弱衍射晶体进行高质量的结构确定.

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.
  • 在X射线晶体学.

背景情况:

  • 在确定蛋白质和核酸等生物分子的原子结构方面,X射线衍射是至关重要的.
  • 确定大型组件 (例如,核糖体) 的结构是具有挑战性的,因为在分辨率低于4 Å的弱衍射.
  • 目前的精制方法对于部分未知组件的宏分子组件失败,需要整个复合体的高分辨率起始模型.

研究的目的:

  • 从低分辨率的X射线衍射数据开发一种新的方法来确定大型生物组件的结构.
  • 为了能够准确地分析弱衍射晶体的结构,克服了传统精炼方法的局限性.
  • 提高宏分子复合体结构模型的质量,其中一些组件是已知的,而另一些组件是未知的.

主要方法:

  • 引入一种方法,将已知同类结构的信息纳入其中.
  • 允许全球和本地对同质模型的变形来解释进化分歧.
  • 使用与自由R因子 (R ((free)) 的交叉验证来优化变形和同质模型的影响.
  • 将该方法应用于具有低分辨率数据 (3.5-5 Å) 和已知的高分辨率结构的测试案例.

主要成果:

  • 与传统改进相比,模型准确性,二次结构定义和电子密度图质量的显著改进.

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Assessing Two-dimensional Crystallization Trials of Small Membrane Proteins for Structural Biology Studies by Electron Crystallography
09:23

Assessing Two-dimensional Crystallization Trials of Small Membrane Proteins for Structural Biology Studies by Electron Crystallography

Published on: October 30, 2010

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  • 来自蛋白质数据库的19个低分辨率晶体结构的再精炼中表现出类似的改进.
  • 从低分辨率衍射数据获得的结构模型质量与高分辨率结构相美.
  • 结论:

    • 开发的方法有效地从低分辨率的X射线衍射数据中确定高质量的结构.
    • 适用于弱衍射晶体,X射线微衍射和新的X射线光源.
    • 同性学信息整合是一种多功能工具,可以超越X射线晶体学,包括冷电子显微镜和先进的光学成像.