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      Convolutional networks for supervised mining of molecular patterns within cellular context

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          Abstract

          Cryo-electron tomograms capture a wealth of structural information on the molecular constituents of cells and tissues. We present DeePiCt (deep picker in context), an open-source deep-learning framework for supervised segmentation and macromolecular complex localization in cryo-electron tomography. To train and benchmark DeePiCt on experimental data, we comprehensively annotated 20 tomograms of Schizosaccharomyces pombe for ribosomes, fatty acid synthases, membranes, nuclear pore complexes, organelles, and cytosol. By comparing DeePiCt to state-of-the-art approaches on this dataset, we show its unique ability to identify low-abundance and low-density complexes. We use DeePiCt to study compositionally distinct subpopulations of cellular ribosomes, with emphasis on their contextual association with mitochondria and the endoplasmic reticulum. Finally, applying pre-trained networks to a HeLa cell tomogram demonstrates that DeePiCt achieves high-quality predictions in unseen datasets from different biological species in a matter of minutes. The comprehensively annotated experimental data and pre-trained networks are provided for immediate use by the community.

          Abstract

          DeePiCt (deep picker in context) is a versatile, open-source deep-learning framework for supervised segmentation and localization of subcellular organelles and biomolecular complexes in cryo-electron tomography.

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          ImageNet classification with deep convolutional neural networks

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            U-Net: Convolutional Networks for Biomedical Image Segmentation

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              New tools for automated high-resolution cryo-EM structure determination in RELION-3

              Here, we describe the third major release of RELION. CPU-based vector acceleration has been added in addition to GPU support, which provides flexibility in use of resources and avoids memory limitations. Reference-free autopicking with Laplacian-of-Gaussian filtering and execution of jobs from python allows non-interactive processing during acquisition, including 2D-classification, de novo model generation and 3D-classification. Per-particle refinement of CTF parameters and correction of estimated beam tilt provides higher resolution reconstructions when particles are at different heights in the ice, and/or coma-free alignment has not been optimal. Ewald sphere curvature correction improves resolution for large particles. We illustrate these developments with publicly available data sets: together with a Bayesian approach to beam-induced motion correction it leads to resolution improvements of 0.2–0.7 Å compared to previous RELION versions.
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                Author and article information

                Contributors
                julia.mahamid@embl.de
                judith.zaugg@embl.de
                Journal
                Nat Methods
                Nat Methods
                Nature Methods
                Nature Publishing Group US (New York )
                1548-7091
                1548-7105
                23 January 2023
                23 January 2023
                2023
                : 20
                : 2
                : 284-294
                Affiliations
                [1 ]GRID grid.4709.a, ISNI 0000 0004 0495 846X, Structural and Computational Biology Unit, European Molecular Biology Laboratory, ; Heidelberg, Germany
                [2 ]GRID grid.7700.0, ISNI 0000 0001 2190 4373, Collaboration for Joint PhD Degree between EMBL and Heidelberg University, Faculty of Biosciences, ; Heidelberg, Germany
                [3 ]GRID grid.7700.0, ISNI 0000 0001 2190 4373, Institute of Pharmacy and Molecular Biotechnology, , Heidelberg University, ; Heidelberg, Germany
                [4 ]GRID grid.4709.a, ISNI 0000 0004 0495 846X, Cell Biology and Biophysics Unit, , European Molecular Biology Laboratory, ; Heidelberg, Germany
                [5 ]GRID grid.4709.a, ISNI 0000 0004 0495 846X, Genome Biology Unit, , European Molecular Biology Laboratory, ; Heidelberg, Germany
                [6 ]Present Address: Computer Science and Artificial Intelligence Lab, ENGIE Lab Crigen, Stains, France
                [7 ]GRID grid.419494.5, ISNI 0000 0001 1018 9466, Present Address: Department of Molecular Sociology, , Max Planck Institute of Biophysics, ; Frankfurt, Germany
                [8 ]Present Address: Santiago GmbH & Co. KG, Willich, Germany
                [9 ]GRID grid.7450.6, ISNI 0000 0001 2364 4210, Present Address: Institute for Computer Science, , Universität Göttingen, ; Göttingen, Germany
                Author information
                http://orcid.org/0000-0002-4691-9501
                http://orcid.org/0000-0002-9903-3667
                http://orcid.org/0000-0003-0901-8701
                http://orcid.org/0000-0002-4327-1068
                http://orcid.org/0000-0003-4388-1349
                http://orcid.org/0000-0001-5333-3640
                http://orcid.org/0000-0001-6562-7187
                http://orcid.org/0000-0002-7397-1321
                http://orcid.org/0000-0003-1334-6388
                http://orcid.org/0000-0001-6968-041X
                http://orcid.org/0000-0001-8324-4040
                Article
                1746
                10.1038/s41592-022-01746-2
                9911354
                36690741
                60d020d0-033d-466f-ad2e-6af617445bc2
                © The Author(s) 2023

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 12 April 2022
                : 2 December 2022
                Funding
                Funded by: FundRef https://doi.org/10.13039/100010665, EC | EU Framework Programme for Research and Innovation H2020 | H2020 Priority Excellent Science | H2020 Marie Skłodowska-Curie Actions (H2020 Excellent Science - Marie Skłodowska-Curie Actions);
                Award ID: 664726
                Award ID: 664726
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/100010663, EC | EU Framework Programme for Research and Innovation H2020 | H2020 Priority Excellent Science | H2020 European Research Council (H2020 Excellent Science - European Research Council);
                Award ID: 760067
                Award Recipient :
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                Article
                Custom metadata
                © The Author(s), under exclusive licence to Springer Nature America, Inc. 2023

                Life sciences
                protein structure predictions,molecular imaging,data mining,image processing
                Life sciences
                protein structure predictions, molecular imaging, data mining, image processing

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