Segmenting multidimensional microscopy data requires high accuracy across many images (e.g., time points or Z slices) and is thus a labor-intensive part of biological image processing pipelines. We present ReSCU-Nets, recurrent convolutional neural networks that use the segmentation results from the previous image in a sequence as a prompt to segment the current image. We demonstrate that ReSCU-Nets outperform state-of-the-art image segmentation models, including nnU-Net and the Segment Anything Model, in different segmentation tasks on time-lapse microscopy sequences. Furthermore, ReSCU-Nets enable human-in-the loop corrections that prevent propagation of segmentation errors throughout image sequences. Using ReSCU-Nets, we investigate the role of gap junctions during Drosophila embryonic wound healing. We show that pharmacological blocking of gap junctions slows down wound closure by disrupting cytoskeletal polarity and cell shape changes necessary to repair the wound. Our results demonstrate that ReSCU-Nets enable the analysis of the molecular and cellular dynamics of tissue morphogenesis from multidimensional microscopy data.
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3 November 2025
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August 11 2025
ReSCU-Nets: Recurrent U-Nets for segmentation of three-dimensional microscopy data
Raymond Hawkins
,
Raymond Hawkins
(Conceptualization, Data curation, Formal analysis, Funding acquisition, Investigation, Methodology, Project administration, Resources, Software, Validation, Visualization, Writing - original draft, Writing - review & editing)
1
Institute of Biomedical Engineering, University of Toronto
, Toronto, Canada
2
Translational Biology and Engineering Program, Ted Rogers Centre for Heart Research, University of Toronto
, Toronto, Canada
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Negar Balaghi
,
Negar Balaghi
(Data curation, Investigation, Validation, Writing - review & editing)
1
Institute of Biomedical Engineering, University of Toronto
, Toronto, Canada
2
Translational Biology and Engineering Program, Ted Rogers Centre for Heart Research, University of Toronto
, Toronto, Canada
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Katheryn E. Rothenberg
,
Katheryn E. Rothenberg
(Data curation, Resources)
1
Institute of Biomedical Engineering, University of Toronto
, Toronto, Canada
2
Translational Biology and Engineering Program, Ted Rogers Centre for Heart Research, University of Toronto
, Toronto, Canada
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Michelle Ly
,
Michelle Ly
(Data curation)
1
Institute of Biomedical Engineering, University of Toronto
, Toronto, Canada
2
Translational Biology and Engineering Program, Ted Rogers Centre for Heart Research, University of Toronto
, Toronto, Canada
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Rodrigo Fernandez-Gonzalez
(Conceptualization, Data curation, Funding acquisition, Methodology, Project administration, Resources, Software, Supervision, Writing - original draft, Writing - review & editing)
1
Institute of Biomedical Engineering, University of Toronto
, Toronto, Canada
2
Translational Biology and Engineering Program, Ted Rogers Centre for Heart Research, University of Toronto
, Toronto, Canada
3Department of Cell and Systems Biology,
University of Toronto
, Toronto, Canada
4
Developmental and Stem Cell Biology Program, The Hospital for Sick Children
, Toronto, Canada
Correspondence to Rodrigo Fernandez-Gonzalez: [email protected]
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Raymond Hawkins
https://orcid.org/0000-0001-8644-6633
Conceptualization, Data curation, Formal analysis, Funding acquisition, Investigation, Methodology, Project administration, Resources, Software, Validation, Visualization, Writing - original draft, Writing - review & editing
1
Institute of Biomedical Engineering, University of Toronto
, Toronto, Canada
2
Translational Biology and Engineering Program, Ted Rogers Centre for Heart Research, University of Toronto
, Toronto, Canada
Negar Balaghi
https://orcid.org/0000-0002-5204-3227
Data curation, Investigation, Validation, Writing - review & editing
1
Institute of Biomedical Engineering, University of Toronto
, Toronto, Canada
2
Translational Biology and Engineering Program, Ted Rogers Centre for Heart Research, University of Toronto
, Toronto, Canada
Katheryn E. Rothenberg
https://orcid.org/0000-0002-8191-7528
Data curation, Resources
1
Institute of Biomedical Engineering, University of Toronto
, Toronto, Canada
2
Translational Biology and Engineering Program, Ted Rogers Centre for Heart Research, University of Toronto
, Toronto, Canada
Michelle Ly
https://orcid.org/0000-0002-6495-8598
Data curation
1
Institute of Biomedical Engineering, University of Toronto
, Toronto, Canada
2
Translational Biology and Engineering Program, Ted Rogers Centre for Heart Research, University of Toronto
, Toronto, Canada
Rodrigo Fernandez-Gonzalez
https://orcid.org/0000-0003-0770-744X
Conceptualization, Data curation, Funding acquisition, Methodology, Project administration, Resources, Software, Supervision, Writing - original draft, Writing - review & editing
1
Institute of Biomedical Engineering, University of Toronto
, Toronto, Canada
2
Translational Biology and Engineering Program, Ted Rogers Centre for Heart Research, University of Toronto
, Toronto, Canada
3Department of Cell and Systems Biology,
University of Toronto
, Toronto, Canada
4
Developmental and Stem Cell Biology Program, The Hospital for Sick Children
, Toronto, Canada
Correspondence to Rodrigo Fernandez-Gonzalez: [email protected]
Disclosures: R. Fernandez-Gonzalez reported nonfinancial support from NVIDIA during the conduct of the study. No other disclosures were reported.
K.E. Rothenberg’s current affiliation is Department of Biology, University of Iowa, Iowa City, IA, USA.
Received:
June 16 2025
Revision Received:
July 22 2025
Accepted:
July 23 2025
Online ISSN: 1540-8140
Print ISSN: 0021-9525
Funding
Funder(s):
Natural Sciences and Engineering Research Council of Canada
- Award Id(s): 418438-13
Funder(s):
Ontario Graduate Scholarship
Funder(s):
Canadian Institutes of Health Research
- Award Id(s): 156279,186188
Funder(s):
Ted Rogers Centre for Heart Research
Funder(s):
Canada Foundation for Innovation
- Award Id(s): 30279
Funder(s):
Canada Research Chair in Quantitative Cell Biology and Morphogenesis
Funder(s):
NVIDIA
© 2025 Hawkins et al.
2025
Hawkins et al.
This article is distributed under the terms as described at https://rupress.org/pages/terms102024/.
J Cell Biol (2025) 224 (11): e202506102.
Article history
Received:
June 16 2025
Revision Received:
July 22 2025
Accepted:
July 23 2025
Citation
Raymond Hawkins, Negar Balaghi, Katheryn E. Rothenberg, Michelle Ly, Rodrigo Fernandez-Gonzalez; ReSCU-Nets: Recurrent U-Nets for segmentation of three-dimensional microscopy data. J Cell Biol 3 November 2025; 224 (11): e202506102. doi: https://doi.org/10.1083/jcb.202506102
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