Open AccessSimplified three-dimensional tissue clearing and incorporation of colorimetric phenotyping
Author(s) -
Kung-Bin 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
Publication year - 2016
Publication title -
scientific reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.24
H-Index - 213
ISSN - 2045-2322
DOI - 10.1038/srep30736
Subject(s) - clearance , light sheet fluorescence microscopy , fluorescence , microscopy , horseradish peroxidase , clearing , computer science , biomedical engineering , fluorescence microscope , chemistry , pathology , optics , medicine , biochemistry , physics , finance , economics , urology , enzyme
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.