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Related Experiment Video

Updated: Mar 14, 2026

Cryo-Electron Tomography Remote Data Collection and Subtomogram Averaging

Cryo-Electron Tomography Remote Data Collection and Subtomogram Averaging

Published on: July 12, 2022

Resolving macromolecular structures from electron cryo-tomography data using subtomogram averaging in RELION.

Tanmay A M Bharat¹, Sjors H W Scheres¹

¹Structural Studies Division, MRC Laboratory of Molecular Biology, Cambridge, UK.

Nature Protocols

|September 30, 2016

Summary

This summary is machine-generated.

This study presents a protocol for subtomogram averaging using RELION software, enabling high-resolution 3D structure determination of macromolecular complexes in situ from electron cryo-tomography data.

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Area of Science:

Structural biology
Biophysics
Microscopy

Background:

Electron cryo-tomography (cryo-ET) generates 3D tomograms of cellular structures.
Subtomogram averaging enhances resolution for in situ macromolecular complex analysis.
This technique is increasingly used for initial model generation in single-particle analysis.

Purpose of the Study:

To describe a protocol for subtomogram averaging using RELION software.
To integrate subtomogram averaging into existing RELION workflows for single-particle analysis.
To demonstrate the application of the protocol for high-resolution structure determination.

Main Methods:

Utilized RELION software for subtomogram averaging of cryo-ET data.
Developed a protocol integrated with existing single-particle analysis workflows.
Calculated 3D contrast transfer function (CTF) models.
Performed classification and refinement on cryo-ET data.

Main Results:

Successfully applied subtomogram averaging to hepatitis B capsid particles and Saccharomyces cerevisiae 80S ribosomes.
Achieved high-resolution 3D maps with resolved secondary structure elements.
Demonstrated the utility of RELION for in situ structural biology.

Conclusions:

The described protocol enables high-resolution structure determination of macromolecular complexes in situ using cryo-ET and RELION.
The integration into existing RELION workflows simplifies the process for users.
The method allows for detailed structural insights, including secondary structure resolution.