{ "abstract": "Vertebrate organs undergo massive growth during the post-embryonic period. Yet, our understanding of how this organ-wide process is organized at single-cell resolution has been limited by an inability to monitor individual cells of different types throughout intact adult organs. Here we establish an integrated workflow of whole adult-organ expansion microscopy (WAO-ExM) that enables in toto single-cell-resolved visualization of every hepatocyte within a complete adult vertebrate liver. Using transgenic reporters to label hepatocyte nuclei, we quantified cell expansion dynamics across the entire post-embryonic growth period, finding that an intact adult liver spanning ~5 mm thickness had an average of 1,265,206 hepatocytes. The data further revealed a non-linear growth regimen in which cell number increased by 538-fold, with a temporally concentrated burst that was not reflective of overall body growth. Integration of lineage tracing with WAO-ExM revealed that cell number increases were driven by a few hepatocytes undergoing drastic clonal expansion at the whole-organ scale. Disruption of extracellular matrix laminins decoupled liver shaping from total cell number increases, suggesting independent regulatory control of tissue architecture and cell proliferation. We also utilized WAO-ExM to monitor diseased livers and other adult organs, including heart and pancreas. Altogether, these findings bridge micrometer-scale cell behaviors with centimeter-scale organ growth, and establish a generalizable platform for adult vertebrate organs to be fully resolved at bona fide single-cell resolution.\n\nVertebrate organs undergo massive growth during the post-embryonic period, but it remains unclear how organs are organised at single-cell resolution. This study develops whole adult-organ expansion microscopy (WAO-ExM) to visualize every single hepatocyte in the zebrafish liver during post-embryonic growth.", "abstract_shown": true, "affiliations": [ "Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan", "Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan", "Nano Science and Technology Program, Taiwan International Graduate Program, Academia Sinica, Taipei, Taiwan", "Department of Engineering and System Science, National Tsing Hua University, Hsinchu, Taiwan", "Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan, Miaoli, Taiwan" ], "article_type": "Research Article", "arxiv": null, "author_total": 13, "authors": [ { "affs": [ 0 ], "name": "Hsiao-Yuh Roan" }, { "affs": [ 1 ], "name": "Xuejiao Tian" }, { "affs": [ 2 ], "name": "Wei-Chen Chu" }, { "affs": [ 3 ], "name": "Chia-Ming Lee" }, { "affs": [ 4 ], "name": "Hsin Chen" }, { "name": "Fiorency Santoso" }, { "name": "Chung-Han Wang" }, { "name": "Yu-Hsiu Liu" }, { "name": "Uday Kumar" }, { "name": "He-Yun Hsiao" }, { "name": "Chiou-Hwa Yuh" }, { "name": "Bi-Chang Chen" }, { "name": "Chen-Hui Chen" } ], "bibtex": "https://research.draconitic.com/paper/6fj77282kzxx6.bib", "cited_by": 0, "doi": "10.1371/journal.pbio.3004007", "fulltext_url": "https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3004007", "id": "6fj77282kzxx6", "issns": [ "1545-7885" ], "landing_url": "https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3004007", "language": null, "licence": "CC-BY-4.0", "licence_label": "CC BY 4.0", "licence_source": "journal", "licence_url": "https://creativecommons.org/licenses/by/4.0/", "metadata_licence": null, "native_subjects": [ [ "openalex", "1307|Zebrafish Biomedical Research Applications" ], [ "plos", "/Biology and life sciences/Anatomy/Cardiovascular anatomy/Heart" ], [ "plos", "/Biology and life sciences/Anatomy/Liver" ], [ "plos", "/Biology and life sciences/Anatomy/Liver/Hepatocytes" ], [ "plos", "/Biology and life sciences/Cell biology/Cellular types/Animal cells/Hepatocytes" ], [ "plos", "/Biology and life sciences/Developmental biology/Life cycles/Larvae" ], [ "plos", "/Biology and life sciences/Molecular biology/Molecular biology techniques/Cloning" ], [ "plos", "/Biology and life sciences/Organisms/Eukaryota/Animals/Vertebrates" ], [ "plos", "/Biology and life sciences/Organisms/Eukaryota/Animals/Vertebrates/Fish/Osteichthyes/Zebrafish" ], [ "plos", "/Biology and life sciences/Zoology/Animals/Vertebrates" ], [ "plos", "/Biology and life sciences/Zoology/Animals/Vertebrates/Fish/Osteichthyes/Zebrafish" ], [ "plos", "/Medicine and health sciences/Anatomy/Cardiovascular anatomy/Heart" ], [ "plos", "/Medicine and health sciences/Anatomy/Liver" ], [ "plos", "/Medicine and health sciences/Anatomy/Liver/Hepatocytes" ], [ "plos", "/Medicine and health sciences/Diagnostic medicine/Diagnostic radiology/Gastrointestinal imaging/Liver and spleen scan" ], [ "plos", "/Medicine and health sciences/Radiology and imaging/Diagnostic radiology/Gastrointestinal imaging/Liver and spleen scan" ], [ "plos", "/Research and analysis methods/Animal studies/Experimental organism systems/Animal models/Zebrafish" ], [ "plos", "/Research and analysis methods/Animal studies/Experimental organism systems/Model organisms/Zebrafish" ], [ "plos", "/Research and analysis methods/Imaging techniques/Diagnostic radiology/Gastrointestinal imaging/Liver and spleen scan" ], [ "plos", "/Research and analysis methods/Model organisms/Zebrafish" ], [ "plos", "/Research and analysis methods/Molecular biology techniques/Cloning" ] ], "openalex": "W7214583772", "pdf_url": "https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.3004007&type=printable", "pmcid": null, "pmid": null, "published": "2026-09-28", "publisher": "Public Library of Science", "ris": "https://research.draconitic.com/paper/6fj77282kzxx6.ris", "sources": [ { "first_seen": "2026-10-30T12:17:37Z", "native_id": "10.1371/journal.pbio.3004007", "source": "plos", "url": "https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3004007" } ], "subjects": [ { "field": "medicine", "origin": "plos:diagnostic radiology", "subfield": "imaging" }, { "field": "medicine", "origin": "plos:anatomy", "subfield": "physiology" } ], "title": "Whole-organ single-cell mapping defines growth dynamics and clonal organization in developing and adult zebrafish livers", "title_text": "Whole-organ single-cell mapping defines growth dynamics and clonal organization in developing and adult zebrafish livers", "type": "peer-reviewed", "updated_at": "2026-10-30T12:17:37Z", "url": "https://research.draconitic.com/paper/6fj77282kzxx6", "venue": "PLOS Biology", "versions": [] }