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      Omnipose: a high-precision morphology-independent solution for bacterial cell segmentation

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          Abstract

          Advances in microscopy hold great promise for allowing quantitative and precise measurement of morphological and molecular phenomena at the single-cell level in bacteria; however, the potential of this approach is ultimately limited by the availability of methods to faithfully segment cells independent of their morphological or optical characteristics. Here, we present Omnipose, a deep neural network image-segmentation algorithm. Unique network outputs such as the gradient of the distance field allow Omnipose to accurately segment cells on which current algorithms, including its predecessor, Cellpose, produce errors. We show that Omnipose achieves unprecedented segmentation performance on mixed bacterial cultures, antibiotic-treated cells and cells of elongated or branched morphology. Furthermore, the benefits of Omnipose extend to non-bacterial subjects, varied imaging modalities and three-dimensional objects. Finally, we demonstrate the utility of Omnipose in the characterization of extreme morphological phenotypes that arise during interbacterial antagonism. Our results distinguish Omnipose as a powerful tool for characterizing diverse and arbitrarily shaped cell types from imaging data.

          Abstract

          Omnipose is a deep neural network algorithm for image segmentation that improves upon existing approaches by solving the challenging problem of accurately segmenting morphologically diverse cells from images acquired with any modality.

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          Mask R-CNN

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            ilastik: interactive machine learning for (bio)image analysis

            We present ilastik, an easy-to-use interactive tool that brings machine-learning-based (bio)image analysis to end users without substantial computational expertise. It contains pre-defined workflows for image segmentation, object classification, counting and tracking. Users adapt the workflows to the problem at hand by interactively providing sparse training annotations for a nonlinear classifier. ilastik can process data in up to five dimensions (3D, time and number of channels). Its computational back end runs operations on-demand wherever possible, allowing for interactive prediction on data larger than RAM. Once the classifiers are trained, ilastik workflows can be applied to new data from the command line without further user interaction. We describe all ilastik workflows in detail, including three case studies and a discussion on the expected performance.
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              Cellpose: a generalist algorithm for cellular segmentation

              Many biological applications require the segmentation of cell bodies, membranes and nuclei from microscopy images. Deep learning has enabled great progress on this problem, but current methods are specialized for images that have large training datasets. Here we introduce a generalist, deep learning-based segmentation method called Cellpose, which can precisely segment cells from a wide range of image types and does not require model retraining or parameter adjustments. Cellpose was trained on a new dataset of highly varied images of cells, containing over 70,000 segmented objects. We also demonstrate a three-dimensional (3D) extension of Cellpose that reuses the two-dimensional (2D) model and does not require 3D-labeled data. To support community contributions to the training data, we developed software for manual labeling and for curation of the automated results. Periodically retraining the model on the community-contributed data will ensure that Cellpose improves constantly.
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                Author and article information

                Contributors
                pwiggins@uw.edu
                mougous@uw.edu
                Journal
                Nat Methods
                Nat Methods
                Nature Methods
                Nature Publishing Group US (New York )
                1548-7091
                1548-7105
                17 October 2022
                17 October 2022
                2022
                : 19
                : 11
                : 1438-1448
                Affiliations
                [1 ]GRID grid.34477.33, ISNI 0000000122986657, Department of Physics, , University of Washington, ; Seattle, WA USA
                [2 ]GRID grid.443970.d, HHMI Janelia Research Campus, ; Ashburn, VA USA
                [3 ]GRID grid.473715.3, ISNI 0000 0004 6475 7299, Centre for Genomic Regulation (CRG), , The Barcelona Institute of Science and Technology, ; Barcelona, Spain
                [4 ]GRID grid.5612.0, ISNI 0000 0001 2172 2676, Universitat Pompeu Fabra (UPF), ; Barcelona, Spain
                [5 ]GRID grid.34477.33, ISNI 0000000122986657, Department of Microbiology, , University of Washington, ; Seattle, WA USA
                [6 ]GRID grid.34477.33, ISNI 0000000122986657, Department of Bioengineering, , University of Washington, ; Seattle, WA USA
                [7 ]GRID grid.34477.33, ISNI 0000000122986657, Howard Hughes Medical Institute, , University of Washington, ; Seattle, WA USA
                Author information
                http://orcid.org/0000-0002-7624-5923
                http://orcid.org/0000-0002-9229-4100
                http://orcid.org/0000-0002-1707-8088
                http://orcid.org/0000-0001-9530-7301
                http://orcid.org/0000-0002-5417-4861
                Article
                1639
                10.1038/s41592-022-01639-4
                9636021
                36253643
                af2845cc-7e54-475a-8f18-36deea408c81
                © The Author(s) 2022

                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
                : 2 December 2021
                : 6 September 2022
                Funding
                Funded by: FundRef https://doi.org/10.13039/100000060, U.S. Department of Health & Human Services | NIH | National Institute of Allergy and Infectious Diseases (NIAID);
                Award ID: AI080609
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/100000011, Howard Hughes Medical Institute (HHMI);
                Funded by: FundRef https://doi.org/10.13039/100000057, U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (NIGMS);
                Award ID: T32-GM008268
                Award ID: GM128191
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/100000002, U.S. Department of Health & Human Services | National Institutes of Health (NIH);
                Award ID: R01-GM128191
                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: 852201
                Award ID: 852201
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/501100011092, University of Chile | Faculty of Physical and Mathematical Sciences | Centro de Excelencia en Geotermia de Los Andes (Andean Geothermal Center of Excellence);
                Categories
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                Custom metadata
                © The Author(s), under exclusive licence to Springer Nature America, Inc. 2022

                Life sciences
                bacteria,imaging
                Life sciences
                bacteria, imaging

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