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Development of a semi-automated brightfield, high-content imaging approach for quantifying morphological effects in zebrafish embryos
Örebro University, School of Science and Technology. (Man-Technology-Environment (MTM) Research Centre - Biology)ORCID iD: 0000-0002-9945-6619
Université Clermont Avergne, Institut Universitaire de Technologie Clermont Auvergne, Clermont-Ferrand, France.
UFZ Helmholtz Centre for Environmental Research, Leipzig, Germany.
Örebro University, School of Science and Technology. (Man-Technology-Environment (MTM) Research Centre - Biology)ORCID iD: 0000-0002-2356-6686
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2026 (English)In: Toxicology Mechanisms and Methods, ISSN 1537-6516, E-ISSN 1537-6524Article in journal (Refereed) Epub ahead of print
Abstract [en]

Environmental pollutants often induce morphological alterations in developing organisms, yet assessments are commonly subjective, limiting reproducibility and sensitivity. We developed and validated a semi-automated brightfield high-content imaging (HCI) pipeline to quantitatively detect morphological changes in zebrafish embryos. Using FishInspector software, we adapted image analysis for microscopy systems without automated embryo positioning, extending applicability across standard laboratory setups.To validate the approach, zebrafish embryos were exposed for 96 hours to two previously characterized pollutant mixtures (PFOS + PCB126; PFOS + B[a]P + arsenate) known to cause developmental effects. The pipeline sensitively quantified phenotypes including reduced swim bladder and shortened body length. These endpoints reflect developmental delay, highlighting the method's ability to capture mechanistically relevant effects. Such changes may reduce physiological performance and behavior, ultimately impacting fish populations.While earlier subjective scoring identified some similar alterations, our findings underscore the advantages of quantitative, semi-automated morphology assessment. The method improves reproducibility, enables standardized comparisons across studies, and increases sensitivity to detecting subtle morphological effects. By integrating brightfield imaging with semi-automated analysis, this approach broadens the toxicological toolbox for developmental hazard assessment and mixture toxicity research.

Place, publisher, year, edition, pages
Taylor & Francis, 2026.
Keywords [en]
FishInspector, High Content Imaging, morphological alterations, zebrafish embryos
National Category
Developmental Biology
Identifiers
URN: urn:nbn:se:oru:diva-127045DOI: 10.1080/15376516.2026.2619427ISI: 001681533200001PubMedID: 41631318OAI: oai:DiVA.org:oru-127045DiVA, id: diva2:2035068
Funder
Knowledge Foundation, 201660019Available from: 2026-02-03 Created: 2026-02-03 Last updated: 2026-02-18Bibliographically approved

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Nilén, GretaKeiter, Steffen H.Scherbak, Nikolai

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