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Automated High-Throughput Light-Sheet Fluorescence Microscopy of Larval Zebrafish

Savannah L. Logan, Christopher Dudley, Ryan P. Baker, Michael J. Taormina, Edouard A. Hay, View ORCID ProfileRaghuveer Parthasarathy
doi: https://doi.org/10.1101/330639
Savannah L. Logan
Materials Science Institute, Institute of Molecular Biology, and Department of Physics, The University of Oregon, Eugene, OR 97403
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Christopher Dudley
Materials Science Institute, Institute of Molecular Biology, and Department of Physics, The University of Oregon, Eugene, OR 97403
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Ryan P. Baker
Materials Science Institute, Institute of Molecular Biology, and Department of Physics, The University of Oregon, Eugene, OR 97403
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Michael J. Taormina
Materials Science Institute, Institute of Molecular Biology, and Department of Physics, The University of Oregon, Eugene, OR 97403
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Edouard A. Hay
Materials Science Institute, Institute of Molecular Biology, and Department of Physics, The University of Oregon, Eugene, OR 97403
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Raghuveer Parthasarathy
Materials Science Institute, Institute of Molecular Biology, and Department of Physics, The University of Oregon, Eugene, OR 97403
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  • ORCID record for Raghuveer Parthasarathy
  • For correspondence: raghu@uoregon.edu
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ABSTRACT

Light sheet fluorescence microscopy enables fast, minimally phototoxic, three-dimensional imaging of live specimens, but is currently limited by low throughput and tedious sample preparation. Here, we describe an automated high-throughput light sheet fluorescence microscope in which specimens are positioned by and imaged within a fluidic system integrated with the sheet excitation and detection optics. We demonstrate the ability of the instrument to rapidly examine live specimens with minimal manual intervention by imaging fluorescent neutrophils over a nearly 0.3 mm3 volume in dozens of larval zebrafish. In addition to revealing considerable inter-individual variability in neutrophil number, known previously from labor-intensive methods, three-dimensional imaging allows assessment of the correlation between the bulk measure of total cellular fluorescence and the spatially resolved measure of actual neutrophil number per animal. We suggest that our simple experimental design should considerably expand the scope and impact of light sheet imaging in the life sciences.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted September 20, 2018.
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Automated High-Throughput Light-Sheet Fluorescence Microscopy of Larval Zebrafish
Savannah L. Logan, Christopher Dudley, Ryan P. Baker, Michael J. Taormina, Edouard A. Hay, Raghuveer Parthasarathy
bioRxiv 330639; doi: https://doi.org/10.1101/330639
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Automated High-Throughput Light-Sheet Fluorescence Microscopy of Larval Zebrafish
Savannah L. Logan, Christopher Dudley, Ryan P. Baker, Michael J. Taormina, Edouard A. Hay, Raghuveer Parthasarathy
bioRxiv 330639; doi: https://doi.org/10.1101/330639

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