Understanding how development is coordinated in multiple tissues and gives rise to fully functional organs or whole organisms necessitates microscopy tools. Over the last decade numerous advances have been made in live-imaging, enabling high resolution imaging of whole organisms at cellular resolution. Yet, these advances mainly rely on mounting the specimen in agarose or aqueous solutions, precluding imaging of organisms whose oxygen uptake depends on ventilation. Here, we implemented a multi-view multi-scale microscopy strategy based on confocal spinning disk microscopy, called Multi-View confocal microScopy (MuViScopy). MuViScopy enables live-imaging of multiple organs with cellular resolution using sample rotation and confocal imaging without the need of sample embedding. We illustrate the capacity of MuViScopy by live-imaging Drosophila melanogaster pupal development throughout metamorphosis, highlighting how internal organs are formed and multiple organ development is coordinated. We foresee that MuViScopy will open the path to better understand developmental processes at the whole organism scale in living systems that necessitates gas exchange by ventilation.
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TECHNIQUES AND RESOURCES ARTICLE|
24 January 2022
Multi-view confocal microscopy enables multiple organ and whole organism live-imaging
Olivier Leroy
,
Olivier Leroy
*
1
Institut Curie, Université PSL, Sorbonne Université, CNRS UMR3215, Inserm U934, Genetics and Developmental Biology, 75005 Paris
, France
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Eric van Leen,
Eric van Leen
*
1
Institut Curie, Université PSL, Sorbonne Université, CNRS UMR3215, Inserm U934, Genetics and Developmental Biology, 75005 Paris
, France
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Philippe Girard
,
Philippe Girard
‡
2
Université de Paris, CNRS, Institut Jacques Monod and Faculty of Basic and Biomedical Sciences, 75006, Paris
, France
‡Authors for correspondence: olivier.renaud@curie.fr; yohanns.bellaiche@curie.fr; philippe.girard@ijm.fr
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Aurélien Villedieu
,
Aurélien Villedieu
1
Institut Curie, Université PSL, Sorbonne Université, CNRS UMR3215, Inserm U934, Genetics and Developmental Biology, 75005 Paris
, France
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Christian Hubert
,
Christian Hubert
3
Errol Laser, Saint Maur des Fossés
, France
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Floris Bosveld
,
Floris Bosveld
1
Institut Curie, Université PSL, Sorbonne Université, CNRS UMR3215, Inserm U934, Genetics and Developmental Biology, 75005 Paris
, France
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Yohanns Bellaïche,
Yohanns Bellaïche
‡
1
Institut Curie, Université PSL, Sorbonne Université, CNRS UMR3215, Inserm U934, Genetics and Developmental Biology, 75005 Paris
, France
‡Authors for correspondence: olivier.renaud@curie.fr; yohanns.bellaiche@curie.fr; philippe.girard@ijm.fr
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Olivier Renaud
Olivier Renaud
‡
1
Institut Curie, Université PSL, Sorbonne Université, CNRS UMR3215, Inserm U934, Genetics and Developmental Biology, 75005 Paris
, France
‡Authors for correspondence: olivier.renaud@curie.fr; yohanns.bellaiche@curie.fr; philippe.girard@ijm.fr
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Olivier Leroy
*
1
Institut Curie, Université PSL, Sorbonne Université, CNRS UMR3215, Inserm U934, Genetics and Developmental Biology, 75005 Paris
, France
Eric van Leen
*
1
Institut Curie, Université PSL, Sorbonne Université, CNRS UMR3215, Inserm U934, Genetics and Developmental Biology, 75005 Paris
, France
Philippe Girard
‡
2
Université de Paris, CNRS, Institut Jacques Monod and Faculty of Basic and Biomedical Sciences, 75006, Paris
, France
Aurélien Villedieu
1
Institut Curie, Université PSL, Sorbonne Université, CNRS UMR3215, Inserm U934, Genetics and Developmental Biology, 75005 Paris
, France
Christian Hubert
3
Errol Laser, Saint Maur des Fossés
, France
Floris Bosveld
1
Institut Curie, Université PSL, Sorbonne Université, CNRS UMR3215, Inserm U934, Genetics and Developmental Biology, 75005 Paris
, France
Yohanns Bellaïche
‡
1
Institut Curie, Université PSL, Sorbonne Université, CNRS UMR3215, Inserm U934, Genetics and Developmental Biology, 75005 Paris
, France
Olivier Renaud
‡
1
Institut Curie, Université PSL, Sorbonne Université, CNRS UMR3215, Inserm U934, Genetics and Developmental Biology, 75005 Paris
, France
*
These authors contributed equally
‡Authors for correspondence: olivier.renaud@curie.fr; yohanns.bellaiche@curie.fr; philippe.girard@ijm.fr
Received:
04 May 2021
Accepted:
13 Jan 2022
Online ISSN: 1477-9129
Print ISSN: 0950-1991
Funding Group:
- Award Group:
- Funder(s): Agence Nationale de la Recherche
- Award Id(s): ANR-10-INBS-04
- Funder(s):
Funding Group:
- Award Group:
- Funder(s): European Research Council
- Award Id(s): 340784
- Funder(s):
Funding Group:
- Award Group:
- Funder(s): Association pour la recherche sur le cancer
- Award Id(s): SL220130607097
- Funder(s):
Funding Group:
- Award Group:
- Funder(s): Marie Skłodowska-Curie Innovative Training Networks
- Award Id(s): 675407
- Funder(s):
Funding Group:
- Award Group:
- Funder(s): Fondation pour la Recherche Médicale
- Award Id(s): FDT201904008163
- Funder(s):
Development dev.199760.
Article history
Received:
04 May 2021
Accepted:
13 Jan 2022
Currently Viewing Accepted Manuscript - Newer Version Available
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Citation
Olivier Leroy, Eric van Leen, Philippe Girard, Aurélien Villedieu, Christian Hubert, Floris Bosveld, Yohanns Bellaïche, Olivier Renaud; Multi-view confocal microscopy enables multiple organ and whole organism live-imaging. Development 2022; dev.199760. doi: https://doi.org/10.1242/dev.199760
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