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Sci Rep. 2016 Aug 8;6:30736. doi: 10.1038/srep30736.

Simplified three-dimensional tissue clearing and incorporation of colorimetric phenotyping.

Author information

1
Department of Bioengineering, University of California, Los Angeles, CA 90095, Los Angeles, USA.
2
Division of Cardiology, Department of Medicine, David Geffen School of Medicine at UCLA, CA 90095, Los Angeles, USA.
3
Department of Molecular, Cellular, and Developmental Biology, University of California, Los Angeles, CA 90095, Los Angeles, USA.
4
Division of Dermatology, Department of Medicine, David Geffen School of Medicine at UCLA, CA 90095, Los Angeles, USA.
5
Department of Neurobiology, David Geffen School of Medicine at UCLA, CA 90095, Los Angeles, USA.
6
Division of Infectious Diseases, Department of Medicine, David Geffen School of Medicine at UCLA, CA 90095, Los Angeles, USA.
7
California NanoSystems Institute, UCLA, CA 90095, Los Angeles, USA.
8
Jonsson Comprehensive Cancer Center, UCLA, CA 90095, Los Angeles, USA.
9
Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research at UCLA, CA 90095, Los Angeles, USA.

Abstract

Tissue clearing methods promise to provide exquisite three-dimensional imaging information; however, there is a need for simplified methods for lower resource settings and for non-fluorescence based phenotyping to enable light microscopic imaging modalities. Here we describe the simplified CLARITY method (SCM) for tissue clearing that preserves epitopes of interest. We imaged the resulting tissues using light sheet microscopy to generate rapid 3D reconstructions of entire tissues and organs. In addition, to enable clearing and 3D tissue imaging with light microscopy methods, we developed a colorimetric, non-fluorescent method for specifically labeling cleared tissues based on horseradish peroxidase conversion of diaminobenzidine to a colored insoluble product. The methods we describe here are portable and can be accomplished at low cost, and can allow light microscopic imaging of cleared tissues, thus enabling tissue clearing and imaging in a wide variety of settings.

PMID:
27498769
PMCID:
PMC4976371
DOI:
10.1038/srep30736
[Indexed for MEDLINE]
Free PMC Article

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