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Updated: Jul 15, 2026

Single Particle Cryo-Electron Microscopy: From Sample to Structure

Single Particle Cryo-Electron Microscopy: From Sample to Structure

Published on: May 29, 2021

Automated image acquisition for single-particle reconstruction using p97 as the biological sample.

I Rouiller¹, J Pulokas, V M Butel

¹Department of Cell Biology, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, California 92037, USA.

Journal of Structural Biology

|July 27, 2001

Summary

The Leginon automated system collects cryo-electron microscopy data for single particles, like the AAA ATPase p97, with no operator intervention. This automation matches experienced microscopists

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Single Particle Cryo-Electron Microscopy: From Sample to Structure

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Area of Science:

Structural Biology
Biophysics
Cryo-Electron Microscopy

Background:

Cryo-electron microscopy (Cryo-EM) is crucial for determining the 3D structure of biological molecules.
Manual data collection in Cryo-EM is time-consuming and requires expert operation.
Automated data acquisition systems aim to improve efficiency and reproducibility.

Purpose of the Study:

To evaluate the performance of the Leginon automated system for single-particle cryo-electron microscopy data collection.
To compare the quality of data acquired by Leginon with data collected manually.
To assess the suitability of Leginon for studying protein structures like AAA ATPase p97.

Main Methods:

Utilized the Leginon automated system for cryo-electron micrograph acquisition of single particles.
Collected data under low-dose conditions with minimal operator intervention.
Acquired images at two different defocus values for each specimen.
Calculated two-dimensional projection maps of the AAA ATPase p97.

Main Results:

Leginon successfully collected high-quality cryo-electron microscopy data for single particles.
Automated data acquisition by Leginon yielded results comparable to manual methods.
The system demonstrated proficiency in acquiring data for the AAA ATPase p97.

Conclusions:

The Leginon system performs comparably to experienced microscopists in single-particle data acquisition.
Automation in cryo-electron microscopy offers significant advantages in efficiency and consistency.
Leginon represents a viable tool for advancing structural biology research through automated data collection.