Improving heterogeneous resolution in cryo-EM volumes reconstructions with pose refinement

Silvera Coeff, Diego

Supervisor(es): Lecumberry, Federico - Bartesaghi, Alberto

Resumen:

Single-particle cryo-electron microscopy (cryo-EM) has emerged as a transformative technique for determining the three-dimensional structures of macromolecular complexes at near-atomic resolution. Its ability to visualize biomolecules in multiple functional states without the need for crystallization has provided unprecedented insights into their structure, dynamics, and mechanisms, making it a cornerstone in structural biology and drug discovery. Despite its success, cryo-EM faces several challenges that limit the achievable resolution and accuracy of reconstructions. Chief among these are the inherently low signal-to-noise ratio (SNR) of raw micrographs, the difficulty in accurately estimating particle orientations (pose estimation), and the presence of conformational and compositional heterogeneity in the sample. In recent years, deep learning has emerged as a leading approach for addressing these limitations, offering powerful methods for denoising, pose refinement, and disentangling structural variability. In this work, a method designed to exploit particle heterogeneity for iterative pose refinement is presented. The approach integrates two state-of-the-art tools : cryoDRGN, which models structural variability using deep generative networks, and Frealign, which performs high-resolution 3D refinement. These tools were combined into a unified pipeline and tested on real cryo-EM datasets, demonstrating the potential of the method to improve both the accuracy of pose estimation and the quality of heterogeneous reconstructions.

Detalles Bibliográficos
2025
Beca de Maestría ANII
Cryo-electron microscopy (cryo-EM)
Heterogeneous 3D reconstruction
Deep generative models
Latent space analysis
UMAP
Clustering
Inglés
Universidad de la República
COLIBRI
https://hdl.handle.net/20.500.12008/53431
Acceso abierto
Licencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0)
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author Silvera Coeff, Diego
author_facet Silvera Coeff, Diego
author_role author
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collection COLIBRI
dc.contributor.filiacion.none.fl_str_mv Silvera Coeff Diego, Universidad de la República (Uruguay). Facultad de Ingeniería.
dc.creator.advisor.none.fl_str_mv Lecumberry, Federico
Bartesaghi, Alberto
dc.creator.none.fl_str_mv Silvera Coeff, Diego
dc.date.accessioned.none.fl_str_mv 2026-02-10T20:42:47Z
dc.date.available.none.fl_str_mv 2026-02-10T20:42:47Z
dc.date.issued.none.fl_str_mv 2025
dc.description.abstract.none.fl_txt_mv Single-particle cryo-electron microscopy (cryo-EM) has emerged as a transformative technique for determining the three-dimensional structures of macromolecular complexes at near-atomic resolution. Its ability to visualize biomolecules in multiple functional states without the need for crystallization has provided unprecedented insights into their structure, dynamics, and mechanisms, making it a cornerstone in structural biology and drug discovery. Despite its success, cryo-EM faces several challenges that limit the achievable resolution and accuracy of reconstructions. Chief among these are the inherently low signal-to-noise ratio (SNR) of raw micrographs, the difficulty in accurately estimating particle orientations (pose estimation), and the presence of conformational and compositional heterogeneity in the sample. In recent years, deep learning has emerged as a leading approach for addressing these limitations, offering powerful methods for denoising, pose refinement, and disentangling structural variability. In this work, a method designed to exploit particle heterogeneity for iterative pose refinement is presented. The approach integrates two state-of-the-art tools : cryoDRGN, which models structural variability using deep generative networks, and Frealign, which performs high-resolution 3D refinement. These tools were combined into a unified pipeline and tested on real cryo-EM datasets, demonstrating the potential of the method to improve both the accuracy of pose estimation and the quality of heterogeneous reconstructions.
dc.description.sponsorship.none.fl_txt_mv Beca de Maestría ANII
dc.format.extent.es.fl_str_mv 116 p.
dc.format.mimetype.es.fl_str_mv application/pdf
dc.identifier.citation.es.fl_str_mv Silvera Coeff, D. Improving heterogeneous resolution in cryo-EM volumes reconstructions with pose refinement [en línea]. Tesis de maestría. Montevideo : Udelar. FI. IIE, 2025.
dc.identifier.issn.none.fl_str_mv 1688-2806
dc.identifier.uri.none.fl_str_mv https://hdl.handle.net/20.500.12008/53431
dc.language.iso.none.fl_str_mv en
eng
dc.publisher.es.fl_str_mv Udelar.FI.
dc.rights.license.none.fl_str_mv Licencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0)
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
dc.source.none.fl_str_mv reponame:COLIBRI
instname:Universidad de la República
instacron:Universidad de la República
dc.subject.es.fl_str_mv Cryo-electron microscopy (cryo-EM)
Heterogeneous 3D reconstruction
Deep generative models
Latent space analysis
UMAP
Clustering
dc.title.none.fl_str_mv Improving heterogeneous resolution in cryo-EM volumes reconstructions with pose refinement
dc.type.es.fl_str_mv Tesis de maestría
dc.type.none.fl_str_mv info:eu-repo/semantics/masterThesis
dc.type.version.none.fl_str_mv info:eu-repo/semantics/acceptedVersion
description Single-particle cryo-electron microscopy (cryo-EM) has emerged as a transformative technique for determining the three-dimensional structures of macromolecular complexes at near-atomic resolution. Its ability to visualize biomolecules in multiple functional states without the need for crystallization has provided unprecedented insights into their structure, dynamics, and mechanisms, making it a cornerstone in structural biology and drug discovery. Despite its success, cryo-EM faces several challenges that limit the achievable resolution and accuracy of reconstructions. Chief among these are the inherently low signal-to-noise ratio (SNR) of raw micrographs, the difficulty in accurately estimating particle orientations (pose estimation), and the presence of conformational and compositional heterogeneity in the sample. In recent years, deep learning has emerged as a leading approach for addressing these limitations, offering powerful methods for denoising, pose refinement, and disentangling structural variability. In this work, a method designed to exploit particle heterogeneity for iterative pose refinement is presented. The approach integrates two state-of-the-art tools : cryoDRGN, which models structural variability using deep generative networks, and Frealign, which performs high-resolution 3D refinement. These tools were combined into a unified pipeline and tested on real cryo-EM datasets, demonstrating the potential of the method to improve both the accuracy of pose estimation and the quality of heterogeneous reconstructions.
eu_rights_str_mv openAccess
format masterThesis
id COLIBRI_61f28bb1edf9537345854ec54b59ec89
identifier_str_mv Silvera Coeff, D. Improving heterogeneous resolution in cryo-EM volumes reconstructions with pose refinement [en línea]. Tesis de maestría. Montevideo : Udelar. FI. IIE, 2025.
1688-2806
instacron_str Universidad de la República
institution Universidad de la República
instname_str Universidad de la República
language eng
language_invalid_str_mv en
network_acronym_str COLIBRI
network_name_str COLIBRI
oai_identifier_str oai:colibri.udelar.edu.uy:20.500.12008/53431
publishDate 2025
reponame_str COLIBRI
repository.mail.fl_str_mv karina.camps@seciu.edu.uy
repository.name.fl_str_mv COLIBRI - Universidad de la República
repository_id_str 4771
rights_invalid_str_mv Licencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0)
spelling Silvera Coeff Diego, Universidad de la República (Uruguay). Facultad de Ingeniería.2026-02-10T20:42:47Z2026-02-10T20:42:47Z2025Silvera Coeff, D. Improving heterogeneous resolution in cryo-EM volumes reconstructions with pose refinement [en línea]. Tesis de maestría. Montevideo : Udelar. FI. IIE, 2025.1688-2806https://hdl.handle.net/20.500.12008/53431Single-particle cryo-electron microscopy (cryo-EM) has emerged as a transformative technique for determining the three-dimensional structures of macromolecular complexes at near-atomic resolution. Its ability to visualize biomolecules in multiple functional states without the need for crystallization has provided unprecedented insights into their structure, dynamics, and mechanisms, making it a cornerstone in structural biology and drug discovery. Despite its success, cryo-EM faces several challenges that limit the achievable resolution and accuracy of reconstructions. Chief among these are the inherently low signal-to-noise ratio (SNR) of raw micrographs, the difficulty in accurately estimating particle orientations (pose estimation), and the presence of conformational and compositional heterogeneity in the sample. In recent years, deep learning has emerged as a leading approach for addressing these limitations, offering powerful methods for denoising, pose refinement, and disentangling structural variability. In this work, a method designed to exploit particle heterogeneity for iterative pose refinement is presented. The approach integrates two state-of-the-art tools : cryoDRGN, which models structural variability using deep generative networks, and Frealign, which performs high-resolution 3D refinement. These tools were combined into a unified pipeline and tested on real cryo-EM datasets, demonstrating the potential of the method to improve both the accuracy of pose estimation and the quality of heterogeneous reconstructions.Submitted by Ribeiro Jorge (jribeiro@fing.edu.uy) on 2026-02-04T16:37:11Z No. of bitstreams: 2 license_rdf: 27293 bytes, checksum: d62648cf14c1e37917d392ac87012955 (MD5) Sil25.pdf: 130891457 bytes, checksum: 65494f16478ac75505ea64843911deab (MD5)Approved for entry into archive by Machado Jimena (jmachado@fing.edu.uy) on 2026-02-06T17:29:36Z (GMT) No. of bitstreams: 2 license_rdf: 27293 bytes, checksum: d62648cf14c1e37917d392ac87012955 (MD5) Sil25.pdf: 130891457 bytes, checksum: 65494f16478ac75505ea64843911deab (MD5)Made available in DSpace by Camps Karina (karina.camps@seciu.edu.uy) on 2026-02-10T20:42:47Z (GMT). No. of bitstreams: 2 license_rdf: 27293 bytes, checksum: d62648cf14c1e37917d392ac87012955 (MD5) Sil25.pdf: 130891457 bytes, checksum: 65494f16478ac75505ea64843911deab (MD5) Previous issue date: 2025Beca de Maestría ANII116 p.application/pdfenengUdelar.FI.Las obras depositadas en el Repositorio se rigen por la Ordenanza de los Derechos de la Propiedad Intelectual de la Universidad de la República.(Res. Nº 91 de C.D.C. de 8/III/1994 – D.O. 7/IV/1994) y por la Ordenanza del Repositorio Abierto de la Universidad de la República (Res. Nº 16 de C.D.C. de 07/10/2014)info:eu-repo/semantics/openAccessLicencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0)Cryo-electron microscopy (cryo-EM)Heterogeneous 3D reconstructionDeep generative modelsLatent space analysisUMAPClusteringImproving heterogeneous resolution in cryo-EM volumes reconstructions with pose refinementTesis de maestríainfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/acceptedVersionreponame:COLIBRIinstname:Universidad de la Repúblicainstacron:Universidad de la RepúblicaSilvera Coeff, DiegoLecumberry, FedericoBartesaghi, AlbertoUniversidad de la República (Uruguay). 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- Universidad de la Repúblicafalse
spellingShingle Improving heterogeneous resolution in cryo-EM volumes reconstructions with pose refinement
Silvera Coeff, Diego
Cryo-electron microscopy (cryo-EM)
Heterogeneous 3D reconstruction
Deep generative models
Latent space analysis
UMAP
Clustering
status_str acceptedVersion
title Improving heterogeneous resolution in cryo-EM volumes reconstructions with pose refinement
title_full Improving heterogeneous resolution in cryo-EM volumes reconstructions with pose refinement
title_fullStr Improving heterogeneous resolution in cryo-EM volumes reconstructions with pose refinement
title_full_unstemmed Improving heterogeneous resolution in cryo-EM volumes reconstructions with pose refinement
title_short Improving heterogeneous resolution in cryo-EM volumes reconstructions with pose refinement
title_sort Improving heterogeneous resolution in cryo-EM volumes reconstructions with pose refinement
topic Cryo-electron microscopy (cryo-EM)
Heterogeneous 3D reconstruction
Deep generative models
Latent space analysis
UMAP
Clustering
url https://hdl.handle.net/20.500.12008/53431