{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T02:09:53Z","timestamp":1783994993003,"version":"3.55.0"},"reference-count":66,"publisher":"Public Library of Science (PLoS)","issue":"12","license":[{"start":{"date-parts":[[2023,12,7]],"date-time":"2023-12-07T00:00:00Z","timestamp":1701907200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100016958","name":"Office of Research Infrastructure Programs, National Institutes of Health","doi-asserted-by":"publisher","award":["R44-OD024879"],"award-info":[{"award-number":["R44-OD024879"]}],"id":[{"id":"10.13039\/100016958","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000066","name":"National Institute of Environmental Health Sciences","doi-asserted-by":"publisher","award":["P42-ES031009"],"award-info":[{"award-number":["P42-ES031009"]}],"id":[{"id":"10.13039\/100000066","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>\n                    Normal development of the immune system is essential for overall health and disease resistance. Bony fish, such as the zebrafish (\n                    <jats:italic>Danio rerio<\/jats:italic>\n                    ), possess all the major immune cell lineages as mammals and can be employed to model human host response to immune challenge. Zebrafish neutrophils, for example, are present in the transparent larvae as early as 48 hours post fertilization and have been examined in numerous infection and immunotoxicology reports. One significant advantage of the zebrafish model is the ability to affordably generate high numbers of individual larvae that can be arrayed in multi-well plates for high throughput genetic and chemical exposure screens. However, traditional workflows for imaging individual larvae have been limited to low-throughput studies using traditional microscopes and manual analyses. Using a newly developed, parallelized microscope, the Multi-Camera Array Microscope (MCAM\u2122), we have optimized a rapid, high-resolution algorithmic method to count fluorescently labeled cells in zebrafish larvae\n                    <jats:italic>in vivo<\/jats:italic>\n                    . Using transgenic zebrafish larvae, in which neutrophils express EGFP, we captured 18 gigapixels of images across a full 96-well plate, in 75 seconds, and processed the resulting datastream, counting individual fluorescent neutrophils in all individual larvae in 5 minutes. This automation is facilitated by a machine learning segmentation algorithm that defines the most in-focus view of each larva in each well after which pixel intensity thresholding and blob detection are employed to locate and count fluorescent cells. We validated this method by comparing algorithmic neutrophil counts to manual counts in larvae subjected to changes in neutrophil numbers, demonstrating the utility of this approach for high-throughput genetic and chemical screens where a change in neutrophil number is an endpoint metric. Using the MCAM\u2122 we have been able to, within minutes, acquire both enough data to create an automated algorithm and execute a biological experiment with statistical significance. Finally, we present this open-source software package which allows the user to train and evaluate a custom machine learning segmentation model and use it to localize zebrafish and analyze cell counts within the segmented region of interest. This software can be modified as needed for studies involving other zebrafish cell lineages using different transgenic reporter lines and can also be adapted for studies using other amenable model species.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0295711","type":"journal-article","created":{"date-parts":[[2023,12,7]],"date-time":"2023-12-07T13:46:21Z","timestamp":1701956781000},"page":"e0295711","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":8,"title":["Automated, high-throughput quantification of EGFP-expressing neutrophils in zebrafish by machine learning and a highly-parallelized microscope"],"prefix":"10.1371","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5431-8634","authenticated-orcid":true,"given":"John","family":"Efromson","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuliano","family":"Ferrero","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6863-2210","authenticated-orcid":true,"given":"Aur\u00e9lien","family":"B\u00e8gue","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-4114-4362","authenticated-orcid":true,"given":"Thomas Jedidiah Jenks","family":"Doman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-6499-9068","authenticated-orcid":true,"given":"Clay","family":"Dugo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andi","family":"Barker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Veton","family":"Saliu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul","family":"Reamey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4557-9525","authenticated-orcid":true,"given":"Kanghyun","family":"Kim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4657-4603","authenticated-orcid":true,"given":"Mark","family":"Harfouche","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6083-1311","authenticated-orcid":true,"given":"Jeffrey A.","family":"Yoder","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2023,12,7]]},"reference":[{"key":"pone.0295711.ref001","doi-asserted-by":"crossref","first-page":"1377","DOI":"10.1016\/j.immuni.2021.06.006","volume":"54","author":"GL Burn","year":"2021","journal-title":"The Neutrophil. Immunity"},{"key":"pone.0295711.ref002","doi-asserted-by":"crossref","first-page":"761","DOI":"10.1038\/ni.2062","article-title":"The junctional adhesion molecule JAM-C regulates polarized transendothelial migration of neutrophils in vivo","volume":"12","author":"A Woodfin","year":"2011","journal-title":"Nat Immunol"},{"key":"pone.0295711.ref003","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1038\/ni.2769","article-title":"Perivascular macrophages mediate neutrophil recruitment during bacterial skin infection","volume":"15","author":"A Abtin","year":"2014","journal-title":"Nat Immunol"},{"key":"pone.0295711.ref004","doi-asserted-by":"crossref","first-page":"3976","DOI":"10.1182\/blood-2006-05-024075","article-title":"A transgenic zebrafish model of neutrophilic inflammation","volume":"108","author":"SA 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Gawish","year":"2022","journal-title":"eLife"},{"key":"pone.0295711.ref008","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1007\/s00441-009-0908-5","article-title":"Neutrophils promote inflammatory angiogenesis via release of preformed VEGF in an in vivo corneal model","volume":"339","author":"Y Gong","year":"2010","journal-title":"Cell Tissue Res"},{"key":"pone.0295711.ref009","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1097\/SHK.0000000000000146","article-title":"Role of neutrophil extracellular traps following injury","volume":"41","author":"F-C Liu","year":"2014","journal-title":"Shock"},{"key":"pone.0295711.ref010","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1016\/0002-9343(90)90187-I","article-title":"A new familial immunodeficiency disorder characterized by severe neutropenia, a defective marrow release mechanism, and hypogammaglobulinemia","volume":"89","author":"M Wetzler","year":"1990","journal-title":"Am J 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unconventional behavior of neutrophils in developing zebrafish","volume":"111","author":"D Le Guyader","year":"2008","journal-title":"Blood"},{"key":"pone.0295711.ref021","doi-asserted-by":"crossref","first-page":"7014","DOI":"10.1182\/blood-2010-12-325712","article-title":"Sumoylation of CCAAT\/enhancer\u2013binding protein \u03b1 promotes the biased primitive hematopoiesis of zebrafish","volume":"117","author":"H Yuan","year":"2011","journal-title":"Blood"},{"key":"pone.0295711.ref022","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1186\/1471-213X-7-42","article-title":"The zebrafish lysozyme C promoter drives myeloid-specific expression in transgenic fish","volume":"7","author":"C Hall","year":"2007","journal-title":"BMC Dev Biol"},{"key":"pone.0295711.ref023","doi-asserted-by":"crossref","first-page":"2390","DOI":"10.3390\/molecules19022390","article-title":"Endotoxin molecule lipopolysaccharide-induced zebrafish inflammation model: a novel screening method for 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Thapa","year":"2021","journal-title":"Bioinform Biol Insights."},{"key":"pone.0295711.ref027","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1002\/bdrc.20216","article-title":"A high-content screening assay in transgenic zebrafish identifies two novel activators of fgf signaling.","volume":"93","author":"M Saydmohammed","year":"2011","journal-title":"Birth Defects Res C Embryo Today"},{"key":"pone.0295711.ref028","doi-asserted-by":"crossref","DOI":"10.1242\/bio.058513","article-title":"A versatile, automated and high-throughput drug screening platform for zebrafish embryos.","volume":"10","author":"A Lubin","year":"2021","journal-title":"Biol Open."},{"key":"pone.0295711.ref029","doi-asserted-by":"crossref","DOI":"10.7554\/eLife.74988","article-title":"Gigapixel imaging with a novel multi-camera array microscope.","volume":"11","author":"E Thomson","year":"2022","journal-title":"Elife"},{"key":"pone.0295711.ref030","article-title":"Imaging across multiple spatial scales with the multi-camera array microscope.","author":"M Harfouche","year":"2023","journal-title":"Optica"},{"key":"pone.0295711.ref031","doi-asserted-by":"crossref","first-page":"100454","DOI":"10.1016\/j.iot.2021.100454","article-title":"Low cost cloud based remote microscopy for biological sciences","volume":"18","author":"PV Baudin","year":"2022","journal-title":"Internet of Things"},{"key":"pone.0295711.ref032","volume-title":"Parallelized computational 3D video microscopy of freely moving organisms at multiple gigapixels per second","author":"KC Zhou","year":"2023"},{"key":"pone.0295711.ref033","first-page":"770","volume-title":"Deep residual learning for image recognition","author":"K He","year":"2015"},{"key":"pone.0295711.ref034","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1145\/3065386","article-title":"ImageNet classification with deep convolutional neural networks.","volume":"60","author":"A Krizhevsky","year":"2017","journal-title":"Commun ACM"},{"key":"pone.0295711.ref035","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1007\/s42979-021-00592-x","article-title":"Machine Learning: Algorithms, Real-World Applications and Research Directions.","volume":"2","author":"IH Sarker","year":"2021","journal-title":"SN Comput Sci."},{"key":"pone.0295711.ref036","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1162\/daed_a_01900","article-title":"A golden decade of deep learning: Computing systems & applications","volume":"151","author":"J. Dean","year":"2022","journal-title":"Daedalus"},{"key":"pone.0295711.ref037","first-page":"1440","article-title":"Species Identification of Caterpillar Eggs by Machine Learning Using a Convolutional Neural Network and Massively Parallelized Microscope.","volume":"12","author":"J Efromson","year":"2022","journal-title":"Collect FAO Agric."},{"key":"pone.0295711.ref038","doi-asserted-by":"crossref","first-page":"104572","DOI":"10.1016\/j.compbiomed.2021.104572","article-title":"COVID-19 cough classification using machine learning and global smartphone recordings","volume":"135","author":"M Pahar","year":"2021","journal-title":"Comput Biol Med"},{"key":"pone.0295711.ref039","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1007\/s10462-009-9112-y","article-title":"Machine learning in digital games: a survey","volume":"29","author":"L Galway","year":"2008","journal-title":"Artificial Intelligence Review"},{"key":"pone.0295711.ref040","first-page":"e148","article-title":"Machine learning techniques for biomedical image segmentation: An overview of technical aspects and introduction to state-of-art applications.","volume":"47","author":"H Seo","year":"2020","journal-title":"Med Phys"},{"key":"pone.0295711.ref041","first-page":"234","volume-title":"Medical Image Computing and Computer-Assisted Intervention\u2013MICCAI 2015","author":"O Ronneberger","year":"2015"},{"key":"pone.0295711.ref042","doi-asserted-by":"crossref","first-page":"106578","DOI":"10.1016\/j.aquatox.2023.106578","article-title":"Bacterial clearance and anti-inflammatory effect of Withaferin A against human pathogen of Staphylococcus aureus in infected zebrafish","volume":"260","author":"R Murugan","year":"2023","journal-title":"Aquat Toxicol"},{"key":"pone.0295711.ref043","doi-asserted-by":"crossref","first-page":"2176953","DOI":"10.1080\/1547691X.2023.2176953","article-title":"Legacy and emerging per- and polyfluoroalkyl substances suppress the neutrophil respiratory burst.","volume":"20","author":"DW Phelps","year":"2023","journal-title":"J Immunotoxicol."},{"key":"pone.0295711.ref044","doi-asserted-by":"crossref","first-page":"108466","DOI":"10.1016\/j.fsi.2022.108466","article-title":"Effects of haloxyfop-p-methyl on the developmental toxicity, neurotoxicity, and immunotoxicity in zebrafish","volume":"132","author":"Y Liu","year":"2023","journal-title":"Fish Shellfish Immunol"},{"key":"pone.0295711.ref045","doi-asserted-by":"crossref","first-page":"106234","DOI":"10.1016\/j.biocel.2022.106234","article-title":"Opportunities presented by zebrafish larval models to study neutrophil function in tissues","volume":"148","author":"H Myllym\u00e4ki","year":"2022","journal-title":"Int J Biochem Cell Biol"},{"key":"pone.0295711.ref046","author":"M. Westerfield","year":"2000","journal-title":"The Zebrafish Book: A Guide for the Laboratory Use of Zebrafish"},{"key":"pone.0295711.ref047","author":"C Nusslein-Volhard","year":"2002"},{"key":"pone.0295711.ref048","doi-asserted-by":"crossref","first-page":"e49","DOI":"10.1182\/blood-2010-10-314120","article-title":"mpeg1 promoter transgenes direct macrophage-lineage expression in zebrafish","volume":"117","author":"F Ellett","year":"2011","journal-title":"Blood"},{"key":"pone.0295711.ref049","doi-asserted-by":"crossref","first-page":"2276","DOI":"10.1145\/3343031.3350535","volume-title":"Proceedings of the 27th ACM International Conference on Multimedia.","author":"A Dutta","year":"2019"},{"key":"pone.0295711.ref050","first-page":"1","article-title":"A survey of loss functions for semantic segmentation","author":"S. Jadon","year":"2020","journal-title":"2020 IEEE Conference on Computational Intelligence in Bioinformatics and Computational Biology (CIBCB)."},{"key":"pone.0295711.ref051","volume-title":"Adam: A Method for Stochastic Optimization","author":"DP Kingma","year":"2014"},{"key":"pone.0295711.ref052","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1007\/978-3-319-50835-1_22","volume-title":"Advances in Visual Computing","author":"MA Rahman","year":"2016"},{"key":"pone.0295711.ref053","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1098\/rspb.1980.0020","article-title":"Theory of edge detection","volume":"207","author":"D Marr","year":"1980","journal-title":"Proc R Soc Lond B Biol Sci"},{"key":"pone.0295711.ref054","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1023\/B:VISI.0000029664.99615.94","article-title":"Distinctive Image Features from Scale-Invariant Keypoints.","volume":"60","author":"DG Lowe","year":"2004","journal-title":"Int J Comput 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compositing for depth of field control"},{"key":"pone.0295711.ref059","doi-asserted-by":"crossref","first-page":"7787","DOI":"10.1364\/OE.27.007787","article-title":"Three-dimensional focal stack imaging in scanning transmission X-ray microscopy with an improved reconstruction algorithm","volume":"27","author":"L Ma","year":"2019","journal-title":"Opt Express"},{"key":"pone.0295711.ref060","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1109\/TCBB.2015.2459685","article-title":"FocusALL: Focal Stacking of Microscopic Images Using Modified Harris Corner Response Measure","volume":"13","author":"MS Sigdel","year":"2016","journal-title":"IEEE\/ACM Trans Comput Biol Bioinform"},{"key":"pone.0295711.ref061","doi-asserted-by":"crossref","DOI":"10.1002\/jbio.201700003","article-title":"Convolutional neural network-based malaria diagnosis from focus stack of blood smear images acquired using custom-built slide scanner","volume":"11","author":"GP Gopakumar","year":"2018","journal-title":"J Biophotonics"},{"key":"pone.0295711.ref062","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1167\/tvst.9.2.11","article-title":"Methodological Challenges of Deep Learning in Optical Coherence Tomography for Retinal Diseases: A Review.","volume":"9","author":"RT Yanagihara","year":"2020","journal-title":"Transl Vis Sci Technol."},{"key":"pone.0295711.ref063","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1049\/htl.2018.5098","article-title":"Machine learning approach of automatic identification and counting of blood cells","volume":"6","author":"MM Alam","year":"2019","journal-title":"Healthc Technol Lett"},{"key":"pone.0295711.ref064","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1016\/j.patcog.2016.07.016","article-title":"Bacterial colony counting with Convolutional Neural Networks in Digital Microbiology Imaging.","volume":"61","author":"A Ferrari","year":"2017","journal-title":"Pattern 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