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Cryo-electron Microscopy

Cryo-electron Microscopy

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...

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Related Experiment Video

Updated: Nov 29, 2025

Cryo-EM and Single-Particle Analysis with Scipion

Cryo-EM and Single-Particle Analysis with Scipion

Published on: May 29, 2021

Reliable cryo-EM resolution estimation with modified Fourier shell correlation.

Pawel A Penczek¹

¹Department of Biochemistry and Molecular Biology, The University of Texas - Houston Medical Center, 6431 Fannin Street, Houston, TX 77030, USA.

|November 19, 2020

Summary

This summary is machine-generated.

A new modified Fourier shell correlation (mFSC) method reduces mask artifacts and estimates resolution uncertainties in cryo-EM. This approach provides reliable, artifact-free results for structural analysis.

Keywords:

cryo-EM modified Fourier shell correlation resolution signal-to-noise ratio three-dimensional structure determination

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Last Updated: Nov 29, 2025

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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

Area of Science:

Structural biology
Biophysics
Computational imaging

Background:

Fourier shell correlation (FSC) is crucial for resolution estimation in cryo-electron microscopy (cryo-EM).
Traditional FSC is susceptible to mask-induced artifacts and lacks uncertainty assessment.
Estimating the number of degrees of freedom (ndf) is challenging, impacting reliability.

Purpose of the Study:

Introduce a modified Fourier shell correlation (mFSC) methodology.
Address mask-induced artifacts and FSC uncertainty estimation.
Improve the reliability of resolution estimation in cryo-EM.

Main Methods:

Reordered FSC computation steps to eliminate mask-induced correlations.
Applied a Gaussian window function to refine reciprocal space regions.
Approximated the number of degrees of freedom by combining window and mask effects.

Main Results:

The modified FSC (mFSC) effectively eliminates mask-induced artifacts.
mFSC provides reliable resolution estimations for single-particle and helical structures.
Developed a novel approach for computing robust local resolutions per voxel or functional subunit.

Conclusions:

The mFSC methodology offers a significant improvement over traditional FSC.
mFSC enables more accurate and reliable structural resolution assessment in cryo-EM.
This method facilitates detailed local resolution analysis of complex biological structures.