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High-Resolution 3D Fluorescent Imaging of Intact Tissues

Danny El-Nachef, Amy M. Martinson, Xiulan Yang, Charles E. Murry, W. Robb MacLellan
doi: https://doi.org/10.1101/855254
Danny El-Nachef
1The Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA
2Department of Medicine, Cardiology, University of Washington, Seattle, WA 98195, USA
3Department of Pathology, University of Washington, Seattle, WA 98109, USA
5Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA
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Amy M. Martinson
1The Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA
3Department of Pathology, University of Washington, Seattle, WA 98109, USA
5Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA
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Xiulan Yang
1The Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA
3Department of Pathology, University of Washington, Seattle, WA 98109, USA
5Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA
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Charles E. Murry
1The Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA
2Department of Medicine, Cardiology, University of Washington, Seattle, WA 98195, USA
3Department of Pathology, University of Washington, Seattle, WA 98109, USA
4Department of Bioengineering, University of Washington, Seattle, WA 98105, USA
5Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA
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W. Robb MacLellan
1The Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA
2Department of Medicine, Cardiology, University of Washington, Seattle, WA 98195, USA
5Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA
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  • For correspondence: WRMacLellan@cardiology.washington.edu
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Abstract

Histological analysis of fluorescently labeled tissues has been a critical tool to understand molecular organization in situ. However, assessing molecular structures within large cells and in the context of human organ anatomy has been challenging because it requires penetration of staining reagents and light deep into opaque tissues, while also conforming to the spatial constraints of high-resolution objective lenses. This methodology article describes optimized sample preparation for sub-micron resolution 3D imaging in human and rodent tissues, yielding imaging depth (>100 µm) and resolution (<0.012 µm3 voxel size) that has previously been limited to whole-mount in vitro organoid systems, embryos, and small model organisms. Confocal images of adult human and rodent organs, including heart, kidney, and liver, were generated for several chemical and antibody stains in cleared tissue sections >100 µm thick. This method can be readily adopted by any lab performing routine histology and takes 3 days from the start of tissue preparation to 3D images.

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Posted November 26, 2019.
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High-Resolution 3D Fluorescent Imaging of Intact Tissues
Danny El-Nachef, Amy M. Martinson, Xiulan Yang, Charles E. Murry, W. Robb MacLellan
bioRxiv 855254; doi: https://doi.org/10.1101/855254
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High-Resolution 3D Fluorescent Imaging of Intact Tissues
Danny El-Nachef, Amy M. Martinson, Xiulan Yang, Charles E. Murry, W. Robb MacLellan
bioRxiv 855254; doi: https://doi.org/10.1101/855254

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