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Aging and Disease    2018, Vol. 9 Issue (2) : 262-272     DOI: 10.14336/AD.2017.0613
Orginal Article |
CLARITY for High-resolution Imaging and Quantification of Vasculature in the Whole Mouse Brain
Lin-Yuan Zhang1,Pan Lin2,Jiaji Pan3,Yuanyuan Ma1,Zhenyu Wei4,Lu Jiang3,Liping Wang1,Yaying Song1,Yongting Wang3,Zhijun Zhang3,Kunlin Jin5,Qian Wang2,*,Guo-Yuan Yang1,3,*
1Department of Neurology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
2Medical Image Computing Lab and
3Neuroscience and Neuroengineering Research Center, Med-X Research Institute and School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China
4Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 201999, China
5Department of Pharmacology and Neuroscience, University of North Texas Health Science Center, TX76107, USA
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Abstract  

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 mm3 per mm3, the normalized vascular length was 0.44 ± 0.04 m per mm3, 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.

Keywords brain      clarity      imaging process      mouse      vasculature     
Corresponding Authors: Qian Wang,Guo-Yuan Yang   
Issue Date: 01 April 2018
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Lin-Yuan Zhang
Pan Lin
Jiaji Pan
Yuanyuan Ma
Zhenyu Wei
Lu Jiang
Liping Wang
Yaying Song
Yongting Wang
Zhijun Zhang
Kunlin Jin
Qian Wang
Guo-Yuan Yang
Cite this article:   
Lin-Yuan Zhang,Pan Lin,Jiaji Pan, et al. CLARITY for High-resolution Imaging and Quantification of Vasculature in the Whole Mouse Brain[J]. A&D, 2018, 9(2): 262-272.
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http://www.aginganddisease.org/EN/10.14336/AD.2017.0613     OR     http://www.aginganddisease.org/EN/Y2018/V9/I2/262
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