Improving heterogeneous resolution in cryo-EM volumes reconstructions with pose refinement
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.
| 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) |
| _version_ | 1875692793651789824 |
|---|---|
| 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 |