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Updated: Oct 26, 2025

Cryo-EM and Single-Particle Analysis with Scipion
Published on: May 29, 2021
Identification of incorrectly oriented particles in cryo-EM single particle analysis
Jeison Méndez1, Edgar Garduño2, José María Carazo3
1Posgrado en Ingeniería Eléctrica, Universidad Nacional Autónoma de México, Cd.Universitaria, C.P.04510, Mexico City, Mexico.
Abstract:
The quality of a 3D map produced by the single-particle analysis method is highly dependent on an accurate assignment of orientations to the many experimental images. However, the problem's complexity implies the presence of several local minima in the optimized goal functions. Consequently, validation methods to confirm the angular assignment are very useful to yield higher-resolution 3D maps. In this work, we present a graph-signal-processing-based methodology that analyzes the correlation landscape as a function of the orientation, an approach allowing the estimation of the assigned orientations' reliability. Using this method, we may identify low-reliability images that probably incorrectly contribute to the final 3D reconstruction.
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