The zebrafish has become a widely used animal model due in large part to its accessibility to and usefulness for high-resolution optical imaging. Although zebrafish research has historically focused mostly on early development, in recent years the fish has increasingly been used to study regeneration, cancer metastasis, behavior, and other processes taking place in juvenile and adult animals. However, imaging of live adult zebrafish is extremely challenging, with survival of adult fish limited to a few tens of minutes using standard imaging methods developed for zebrafish embryos and larvae. Here, we describe a new method for imaging intubated adult zebrafish using a specially designed 3D printed chamber for long-term imaging of adult zebrafish on inverted microscope systems. We demonstrate the utility of this new system by nearly day-long observation of neutrophil recruitment to a wound area in living double-transgenic adult casper zebrafish with fluorescently labeled neutrophils and lymphatic vessels, as well as intubating and imaging the same fish repeatedly. We also show that Mexican Cavefish can be intubated and imaged in the same way, showing this method can be used for long-term imaging of adult animals from diverse aquatic species.
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10 February 2022
Long-Term Imaging of Living Adult Zebrafish
Daniel Castranova,
Daniel Castranova
Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD
, USA
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Bakary Samasa,
Bakary Samasa
Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD
, USA
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Marina Venero Galanternik,
Marina Venero Galanternik
Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD
, USA
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Aniket V. Gore,
Aniket V. Gore
Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD
, USA
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Allison E. Goldstein,
Allison E. Goldstein
Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD
, USA
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Jong S. Park,
Jong S. Park
Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD
, USA
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Brant M. Weinstein
Brant M. Weinstein
*
Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD
, USA
*Author for correspondence: WeinsteB@mail.nih.gov
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Daniel Castranova
Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD
, USA
Bakary Samasa
Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD
, USA
Marina Venero Galanternik
Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD
, USA
Aniket V. Gore
Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD
, USA
Allison E. Goldstein
Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD
, USA
Jong S. Park
Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD
, USA
Brant M. Weinstein
*
Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD
, USA
*Author for correspondence: WeinsteB@mail.nih.gov
Received:
30 Mar 2021
Accepted:
30 Nov 2021
Online ISSN: 1477-9129
Print ISSN: 0950-1991
Funding Group:
- Award Group:
- Funder(s): National Institutes of Health
- Award Id(s): ZIA-HD008915
- Funder(s):
Development dev.199667.
Article history
Received:
30 Mar 2021
Accepted:
30 Nov 2021
Currently Viewing Accepted Manuscript - Newer Version Available
21 Feb 2022
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Citation
Daniel Castranova, Bakary Samasa, Marina Venero Galanternik, Aniket V. Gore, Allison E. Goldstein, Jong S. Park, Brant M. Weinstein; Long-Term Imaging of Living Adult Zebrafish. Development 2022; dev.199667. doi: https://doi.org/10.1242/dev.199667
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