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Related Experiment Video

Updated: Nov 29, 2025

Routine Collection of High-Resolution cryo-EM Datasets Using 200 KV Transmission Electron Microscope

Routine Collection of High-Resolution cryo-EM Datasets Using 200 KV Transmission Electron Microscope

Published on: March 16, 2022

High-resolution cryo-EM using beam-image shift at 200 keV.

Jennifer N Cash¹, Sarah Kearns¹, Yilai Li¹

¹Life Sciences Institute, Department of Biological Chemistry, University of Michigan, Ann Arbor, MI 48109, USA.

|November 19, 2020

Summary

Beam-image shift in cryo-electron microscopy (cryo-EM) can be used on 200 keV instruments. Software correction of aberrations significantly improves resolution, enabling high-quality structural analysis.

Keywords:

RELION aldolase single-particle cryo-EM

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Last Updated: Nov 29, 2025

Routine Collection of High-Resolution cryo-EM Datasets Using 200 KV Transmission Electron Microscope

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Published on: March 16, 2022

Single-Particle Cryo-EM Data Collection with Stage Tilt using Leginon

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Published on: July 1, 2022

Strategies for Optimization of Cryogenic Electron Tomography Data Acquisition

Strategies for Optimization of Cryogenic Electron Tomography Data Acquisition

Published on: March 19, 2021

Area of Science:

Structural Biology
Biophysics
Microscopy

Background:

Single-particle cryo-electron microscopy (cryo-EM) uses beam-image shift to enhance data collection throughput.
The impact of beam-image shift on data quality at 200 keV cryo-EM instruments is not well understood.
Computational correction of aberrations in cryo-EM data is an active area of research.

Purpose of the Study:

To evaluate the effect of beam-image shift on cryo-EM data quality at 200 keV.
To determine the extent to which computational methods can correct for aberrations introduced by beam-image shift.
To assess the achievable resolution for cryo-EM data collected using beam-image shift at 200 keV.

Main Methods:

Collected cryo-EM data for aldolase at 200 keV utilizing beam-image shift.
Analyzed the initial data quality, identifying limitations due to instrument beam tilt and particle motion.
Applied particle polishing and iterative aberration correction using RELION software.

Main Results:

Initial data resolution was limited to 4.9 Å due to instrument beam tilt and particle motion.
Post-processing, including particle polishing and aberration correction, significantly improved data quality.
Achieved a final resolution of 2.8 Å for the aldolase structure.

Conclusions:

Beam-image shift data collection is feasible and beneficial on 200 keV cryo-EM instruments.
Computational correction of microscope aberrations is effective in mitigating resolution loss.
This study demonstrates the potential for high-resolution cryo-EM structural determination at 200 keV using advanced data processing techniques.