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      High-throughput phenotyping of nematode cysts

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          Abstract

          The beet cyst nematode Heterodera schachtii is a plant pest responsible for crop loss on a global scale. Here, we introduce a high-throughput system based on computer vision that allows quantifying beet cyst nematode infestation and measuring phenotypic traits of cysts. After recording microscopic images of soil sample extracts in a standardized setting, an instance segmentation algorithm serves to detect nematode cysts in these images. In an evaluation using both ground truth samples with known cyst numbers and manually annotated images, the computer vision approach produced accurate nematode cyst counts, as well as accurate cyst segmentations. Based on such segmentations, cyst features could be computed that served to reveal phenotypical differences between nematode populations in different soils and in populations observed before and after the sugar beet planting period. The computer vision approach enables not only fast and precise cyst counting, but also phenotyping of cyst features under different conditions, providing the basis for high-throughput applications in agriculture and plant breeding research. Source code and annotated image data sets are freely available for scientific use.

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          Most cited references16

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          YOLOv4: Optimal Speed and Accuracy of Object Detection

          There are a huge number of features which are said to improve Convolutional Neural Network (CNN) accuracy. Practical testing of combinations of such features on large datasets, and theoretical justification of the result, is required. Some features operate on certain models exclusively and for certain problems exclusively, or only for small-scale datasets; while some features, such as batch-normalization and residual-connections, are applicable to the majority of models, tasks, and datasets. We assume that such universal features include Weighted-Residual-Connections (WRC), Cross-Stage-Partial-connections (CSP), Cross mini-Batch Normalization (CmBN), Self-adversarial-training (SAT) and Mish-activation. We use new features: WRC, CSP, CmBN, SAT, Mish activation, Mosaic data augmentation, CmBN, DropBlock regularization, and CIoU loss, and combine some of them to achieve state-of-the-art results: 43.5% AP (65.7% AP50) for the MS COCO dataset at a realtime speed of ~65 FPS on Tesla V100. Source code is at https://github.com/AlexeyAB/darknet
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            A Dataset and a Technique for Generalized Nuclear Segmentation for Computational Pathology.

            Nuclear segmentation in digital microscopic tissue images can enable extraction of high-quality features for nuclear morphometrics and other analysis in computational pathology. Conventional image processing techniques, such as Otsu thresholding and watershed segmentation, do not work effectively on challenging cases, such as chromatin-sparse and crowded nuclei. In contrast, machine learning-based segmentation can generalize across various nuclear appearances. However, training machine learning algorithms requires data sets of images, in which a vast number of nuclei have been annotated. Publicly accessible and annotated data sets, along with widely agreed upon metrics to compare techniques, have catalyzed tremendous innovation and progress on other image classification problems, particularly in object recognition. Inspired by their success, we introduce a large publicly accessible data set of hematoxylin and eosin (H&E)-stained tissue images with more than 21000 painstakingly annotated nuclear boundaries, whose quality was validated by a medical doctor. Because our data set is taken from multiple hospitals and includes a diversity of nuclear appearances from several patients, disease states, and organs, techniques trained on it are likely to generalize well and work right out-of-the-box on other H&E-stained images. We also propose a new metric to evaluate nuclear segmentation results that penalizes object- and pixel-level errors in a unified manner, unlike previous metrics that penalize only one type of error. We also propose a segmentation technique based on deep learning that lays a special emphasis on identifying the nuclear boundaries, including those between the touching or overlapping nuclei, and works well on a diverse set of test images.
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              K-Sample Anderson–Darling Tests

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                Author and article information

                Contributors
                Journal
                Front Plant Sci
                Front Plant Sci
                Front. Plant Sci.
                Frontiers in Plant Science
                Frontiers Media S.A.
                1664-462X
                14 September 2022
                2022
                : 13
                : 965254
                Affiliations
                [1] 1Institute of Imaging and Computer Vision (LfB), RWTH Aachen University , Aachen, Germany
                [2] 2Federal Research Center for Cultivated Plants, Julius Kühn Institute (JKI) , Elsdorf, Germany
                [3] 3LemnaTec GmbH , Aachen, Germany
                Author notes

                Edited by: Jianfeng Zhou, University of Missouri, United States

                Reviewed by: Chongyuan Zhang, Purdue University, United States; Rasha Haj Nuaima, Julius Kühn-Institute, Germany; Krzysztof Wieczorek, University of Natural Resources and Life Sciences Vienna, Austria

                *Correspondence: Dorit Merhof dorit.merhof@ 123456lfb.rwth-aachen.de

                This article was submitted to Technical Advances in Plant Science, a section of the journal Frontiers in Plant Science

                †These authors have contributed equally to this work

                Article
                10.3389/fpls.2022.965254
                9515587
                36186075
                16983476-7c19-45e2-92ed-43ff060426e8
                Copyright © 2022 Chen, Daub, Luigs, Jansen, Strauch and Merhof.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 09 June 2022
                : 08 August 2022
                Page count
                Figures: 4, Tables: 1, Equations: 6, References: 16, Pages: 12, Words: 7473
                Funding
                Funded by: Bundesministerium für Bildung und Forschung, doi 10.13039/501100002347;
                Categories
                Plant Science
                Original Research

                Plant science & Botany
                phenotyping,nematode cyst,heterodera schachtii,nematode infestation,sugar beet,cnn

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