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Quantifiable Intravital Light Sheet Microscopy

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Fluorescent Microscopy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2440))

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Abstract

Live imaging of zebrafish embryos that maintains normal development can be difficult to achieve due to a combination of sample mounting, immobilization, and phototoxicity issues that, once overcome, often still results in image quality sufficiently poor that computer-aided analysis or even manual analysis is not possible. Here, we describe our mounting strategy for imaging the zebrafish midbrain–hindbrain boundary (MHB) with light sheet fluorescence microscopy (LSFM) and pilot experiments to create a study-specific set of parameters for semiautomatically tracking cellular movements in the embryonic midbrain primordium during zebrafish segmentation.

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Acknowledgments

We thank Dr. Bruce Riley, Dr. Jennifer Dong, and Dr. Jo-Ann Fleming for care and maintenance of zebrafish colonies and facilities, and we thank Texas A&M University’s Microscopy and Imaging Center for access to the Zeiss Z.1 LSFM. The plasmid containing citrine:H2b was a gift from S.Megason. This work was supported by the Silicon Valley Community Foundation (CZI Imaging Scientist Program, 2019-198168) and the National Institutes of Health (R01 NS088564 and R21 NS109504).

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Authors and Affiliations

  1. Department of Biomedical Engineering, Texas A&M University, College Station, TX, USA

    Holly C. Gibbs, Sreeja Sarasamma, Oscar R. Benavides, Nathan A. Hart, Alvin T. Yeh & Kristen C. Maitland

  2. Microscopy and Imaging Center, Texas A&M University, College Station, TX, USA

    Holly C. Gibbs & Kristen C. Maitland

  3. Department of Cell and Systems Biology, University of Toronto, Toronto, Canada

    David G. Green

  4. Department of Biology and Biochemistry, University of Houston, Houston, TX, USA

    Arne C. Lekven

Authors
  1. Holly C. Gibbs
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  2. Sreeja Sarasamma
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  3. Oscar R. Benavides
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  4. David G. Green
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  5. Nathan A. Hart
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  6. Alvin T. Yeh
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  7. Kristen C. Maitland
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  8. Arne C. Lekven
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Corresponding author

Correspondence to Holly C. Gibbs .

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Editors and Affiliations

  1. Department of Microbiology and Immunology, Center for Human Immunology, The University of Western Ontario, London, ON, Canada

    Bryan Heit

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Gibbs, H.C. et al. (2022). Quantifiable Intravital Light Sheet Microscopy. In: Heit, B. (eds) Fluorescent Microscopy. Methods in Molecular Biology, vol 2440. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2051-9_11

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  • DOI: https://doi.org/10.1007/978-1-0716-2051-9_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2050-2

  • Online ISBN: 978-1-0716-2051-9

  • eBook Packages: Springer Protocols

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