相关实验视频

Updated: Jun 22, 2025

User-friendly, High-throughput, and Fully Automated Data Acquisition Software for Single-particle Cryo-electron Microscopy

User-friendly, High-throughput, and Fully Automated Data Acquisition Software for Single-particle Cryo-electron Microscopy

Published on: July 29, 2021

在100-200 keV的高分辨率单粒子成像使用加坦阿尔卑斯的直接电子探测器

Lieza M Chan¹, Brandon J Courteau¹, Allison Maker¹

¹Department of Cellular and Molecular Pharmacology, University of California, San Francisco, CA 94158, United States.

Journal of structural biology

|June 29, 2024

概括

新的Gatan Alpine探测器显著提高了在较低电压下电子检测的效率,使较小的生物样本能够进行高分辨率的冷电子显微镜 (cryo-EM).

科学领域:

结构生物学结构生物学
生物物理学的生物物理.
材料科学材料科学材料科学

背景情况:

直接电子探测器对于高分辨率的冷电子显微镜 (cryo-EM) 是至关重要的.
电流探测器优化为200-300 keV,但较低的能量 (例如100 keV) 提供了理论上的优势.
针对较低keV优化的探测器可能会增强冷EM功能.

研究的目的:

评估Gatan Alpine探测器的性能,该探测器设计用于100和200keV的操作.
将Alpine探测器的性能与Gatan K3探测器进行比较.
评估Alpine在各种加速电压下对生物样本的高分辨率冷电磁图的实用性.

主要方法:

在100,120和200keV测试了加坦阿尔卑斯探测器.
在Apoferritin和一个不对称的蛋白质复合体上进行单颗粒冷EM实验.
使用ThermoFisher Scientific (TFS) Glacios和Titan Krios显微镜获得的数据.
使用侦探量子效率 (DQE) 测量分析了探测器性能.

主要成果:

与K3.3相比,Alpine在100keV时 (在Nyquist) 显示了约4倍的DQE改善.
在120和200keV时实现了apopherritin在2 Å以上的分辨率重建.
获得了115kDa蛋白质复合体的~3.2 Å分辨率重建.
在格拉西奥斯120keV的阿尔卑斯山重建与克里奥斯/GIF/K3.3的300keV数据相美.
在低端系统上,在100 keV的电压下,对阿波费里丁和阿尔多酶的显示分辨率为 ~ 3 Å.

结论:

加坦阿尔卑斯探测器在较低的加速电压下提供显著的DQE优势.
阿尔派能在较小的粒子和复杂的样品上实现高分辨率的冷电磁波,即使在较不先进的系统上也是如此.
阿尔卑斯可以释放低电压成像的好处,扩大了冷EM的范围.

关键词:

100 keV成像技术使用显微镜硬件的进步显微镜硬件的进步直接探测器是一种直接探测器.加坦阿尔卑斯山的加坦一个LKB1复合体.单颗粒冷电磁器 (Cryo-EM) 是一种

更多相关视频

High-resolution Single Particle Analysis from Electron Cryo-microscopy Images Using SPHIRE

High-resolution Single Particle Analysis from Electron Cryo-microscopy Images Using SPHIRE

Published on: May 16, 2017

Quantitative Atomic-Site Analysis of Functional Dopants/Point Defects in Crystalline Materials by Electron-Channeling-Enhanced Microanalysis

Quantitative Atomic-Site Analysis of Functional Dopants/Point Defects in Crystalline Materials by Electron-Channeling-Enhanced Microanalysis

Published on: May 10, 2021

相关概念视频

Electron Microscope Tomography and Single-particle Reconstruction

Electron Microscope Tomography and Single-particle Reconstruction

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...

Scanning Electron Microscopy

Scanning Electron Microscopy

A scanning electron microscope (SEM) is used to study the surface features of a sample by using an electron beam that scans the sample surface in a two-dimensional manner. Typically, areas between ~1 centimeter to 5 micrometers in width can be imaged. SEM can be used to image bacteria, viruses, tissues as well as larger samples like insects. Conventional SEM gives a magnification ranging from 20X to 30,000X and spatial resolution of 50 to 100 nanometers.
Fundamental Principles
Accelerated...

Transmission Electron Microscopy

Transmission Electron Microscopy

In 1931, physicist Ernst Ruska—building on the idea that magnetic fields can direct an electron beam just as lenses can direct a beam of light in an optical microscope—developed the first prototype of the electron microscope. This development led to the development of the field of electron microscopy. In the transmission electron microscope (TEM), electrons are produced by a hot tungsten element and accelerated by a potential difference in an electron gun, which gives them up to 400...