{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T13:11:21Z","timestamp":1790255481506,"version":"4.1.0"},"reference-count":21,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["MicrobiologyOpen"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Agar plate photography is widely used to document microbial growth, but images captured under routine laboratory lighting are often affected by glare, reflections, shadows, and uneven illumination, which may compromise both visual interpretation and computational analysis. This proof\u2010of\u2010concept study developed and evaluated a low\u2010cost, three\u2010dimensional (3D)\u2010printed lightbox for standardized smartphone\u2010based agar plate imaging. Image characteristics were assessed across five uncultured agar media under traditional and lightbox conditions using Fiji\/ImageJ. Machine learning\u2013assisted analysis was then performed on cultured horse blood agar plates using QuPath. Separate Random Trees pixel classifiers were developed for traditional and lightbox images to distinguish colonies from background agar and noncolony artifacts. A semiautomated QuPath\u2013assisted workflow was also compared with manual colony counts across 14 growth\u2010positive horse blood agar plates. Lightbox imaging reduced reflective artifact, improved illumination consistency, and enhanced visualization of colony margins and hemolysis\u2010associated changes. Traditional\u2010lighting images produced more artifact\u2010related classifier detections, including glare, reflections, and uncolonized agar being detected as colony\u2010like regions. In the exploratory colony\u2010count comparison, semiautomated QuPath\u2013assisted counts showed close agreement with manual reference counts (Pearson\n                    <jats:italic>r<\/jats:italic>\n                    \u2009=\u20090.999; mean absolute error\u2009=\u20092.4 colonies per plate). These findings demonstrate that a low\u2010cost, 3D\u2010printed lightbox can improve the quality and standardization of smartphone\u2010based agar plate images and support more interpretable machine learning\u2013assisted analysis. Further validation using larger independent data sets, additional organisms, media types, and imaging devices is required.\n                  <\/jats:p>","DOI":"10.1002\/mbo3.70399","type":"journal-article","created":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T13:42:21Z","timestamp":1789998141000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Improving Agar Plate Image Quality Using a Low\u2010Cost 3D\u2010Printed Lightbox: A Proof\u2010of\u2010Concept Evaluation for Machine Learning\u2013Assisted Analysis"],"prefix":"10.1002","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-0938-4720","authenticated-orcid":false,"given":"Liam A.","family":"O'Callaghan","sequence":"first","affiliation":[{"name":"Faculty of Health Sciences and Medicine Bond University Gold Coast Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0974-524X","authenticated-orcid":false,"given":"Matthew","family":"Olsen","sequence":"additional","affiliation":[{"name":"Faculty of Health Sciences and Medicine Bond University Gold Coast Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caleb","family":"Kam","sequence":"additional","affiliation":[{"name":"Faculty of Health Sciences and Medicine Bond University Gold Coast Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew","family":"Linnanne","sequence":"additional","affiliation":[{"name":"School of Allied Health, Sport and Social Work Griffith University Nathan Queensland Australia"},{"name":"Physiotherapy Department The Prince Charles Hospital Chermside Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7628-5725","authenticated-orcid":false,"given":"Adrian","family":"Goldsworthy","sequence":"additional","affiliation":[{"name":"Critical Care Research Group The Prince Charles Hospital Chermside Queensland Australia"},{"name":"Institute for Molecular Bioscience The University of Queensland Brisbane Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,21]]},"reference":[{"key":"e_1_2_11_2_1","volume-title":"Recommended Curriculum Guidelines for Undergraduate Microbiology Education","author":"American Society for Microbiology","year":"2024"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.1002175"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-17204-5"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1128\/jmbe.00126-24"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0033695"},{"key":"e_1_2_11_7_1","volume-title":"Blood Agar Plates and Hemolysis Protocols","author":"Buxton R.","year":"2016"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0054072"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.04209-23"},{"key":"e_1_2_11_10_1","volume-title":"Scan 1200: HD Automatic Colony Counter and Inhibition Zone Reader","author":"Interscience","year":"2026"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.01651-23"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-24916-9"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-022-23990-4"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.4103\/2153-3539.157789"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmpb.2017.12.017"},{"key":"e_1_2_11_16_1","volume-title":"schuett colony Quant: Automated Colony Counter System","author":"Schuett\u2010biotec 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