Simplified three-dimensional tissue clearing and incorporation of colorimetric phenotyping

Kevin Sung, Yichen Ding, Jianguo Ma, Harrison Chen, Vincent Huang, Michelle Cheng, Cindy F. Yang, Jocelyn T. Kim, Daniel Eguchi, Dino Di Carlo, Tzung K. Hsiai, Atsushi Nakano, Rajan P. Kulkarni

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

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.

Original languageEnglish (US)
Article number30736
JournalScientific Reports
Volume6
DOIs
StatePublished - Aug 8 2016
Externally publishedYes

ASJC Scopus subject areas

  • General

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