Open AccessCLARITY for High-resolution Imaging and Quantification of Vasculature in the Whole Mouse Brain
Author(s) -
Lin Yuan Zhang,
Pang Lin,
Jiaji Pan,
Yuanyuan Ma,
Zhenyu Wei,
Lu Jiang,
Liping Wang,
Yaying Song,
Yongting Wang,
Zhijun Zhang,
Kunlin Jin,
Qian Wang,
Guo Yuan Yang
Publication year - 2018
Publication title -
aging and disease
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.808
H-Index - 54
ISSN - 2152-5250
DOI - 10.14336/ad.2017.0613
Subject(s) - medicine , clarity , visualization , nuclear medicine , biomedical engineering , pathology , computer science , biology , artificial intelligence , biochemistry
Elucidating the normal structure and distribution of cerebral vascular system is fundamental for understanding its function. However, studies on visualization and whole-brain quantification of vasculature with cellular resolution are limited. Here, we explored the structure of vasculature at the whole-brain level using the newly developed CLARITY technique. Adult male C57BL/6J mice undergoing transient middle cerebral artery occlusion and Tie2-RFP transgenic mice were used. Whole mouse brains were extracted for CLARITY processing. Immunostaining was performed to label vessels. Customized MATLAB code was used for image processing and quantification. Three-dimensional images were visualized using the Vaa3D software. Our results showed that whole mouse brain became transparent using the CLARITY method. Three-dimensional imaging and visualization of vasculature were achieved at the whole-brain level with a 1-μm voxel resolution. The quantitative results showed that the fractional vascular volume was 0.018 ± 0.004 mm 3 per mm 3 , the normalized vascular length was 0.44 ± 0.04 m per mm 3 , and the mean diameter of the microvessels was 4.25 ± 0.08 μm. Furthermore, a decrease in the fractional vascular volume and a decrease in the normalized vascular length were found in the penumbra of ischemic mice compared to controls ( p < 0.05). In conclusion, CLARITY provides a novel approach for mapping vasculature in the whole mouse brain at cellular resolution. CLARITY-optimized algorithms facilitate the assessment of structural change in vasculature after brain injury.