Whole-organ single-cell mapping defines growth dynamics and clonal organization in developing and adult zebrafish livers
5 affiliations
- 1Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan
- 2Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan
- 3Nano Science and Technology Program, Taiwan International Graduate Program, Academia Sinica, Taipei, Taiwan
- 4Department of Engineering and System Science, National Tsing Hua University, Hsinchu, Taiwan
- 5Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan, Miaoli, Taiwan
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.
Vertebrate 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.
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BibTeX
@article{roan2026wholeorgan,
title = {{Whole-organ single-cell mapping defines growth dynamics and clonal organization in developing and adult zebrafish livers}},
author = {Hsiao-Yuh Roan and Xuejiao Tian and Wei-Chen Chu and Chia-Ming Lee and Hsin Chen and Fiorency Santoso and Chung-Han Wang and Yu-Hsiu Liu and Uday Kumar and He-Yun Hsiao and Chiou-Hwa Yuh and Bi-Chang Chen and Chen-Hui Chen},
journal = {PLOS Biology},
year = {2026},
month = sep,
doi = {10.1371/journal.pbio.3004007},
url = {https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3004007},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Whole-organ single-cell mapping defines growth dynamics and clonal organization in developing and adult zebrafish livers
AU - Hsiao-Yuh Roan
AU - Xuejiao Tian
AU - Wei-Chen Chu
AU - Chia-Ming Lee
AU - Hsin Chen
AU - Fiorency Santoso
AU - Chung-Han Wang
AU - Yu-Hsiu Liu
AU - Uday Kumar
AU - He-Yun Hsiao
AU - Chiou-Hwa Yuh
AU - Bi-Chang Chen
AU - Chen-Hui Chen
PY - 2026
DA - 2026/09/28
JO - PLOS Biology
DO - 10.1371/journal.pbio.3004007
UR - https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3004007
AB - 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. Vertebrate 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.
SN - 1545-7885
ER -