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Merck

Tissue clearing of both hard and soft tissue organs with the PEGASOS method.

Cell research (2018-05-31)
Dian Jing, Shiwen Zhang, Wenjing Luo, Xiaofei Gao, Yi Men, Chi Ma, Xiaohua Liu, Yating Yi, Abhijit Bugde, Bo O Zhou, Zhihe Zhao, Quan Yuan, Jian Q Feng, Liang Gao, Woo-Ping Ge, Hu Zhao
要旨

Tissue clearing technique enables visualization of opaque organs and tissues in 3-dimensions (3-D) by turning tissue transparent. Current tissue clearing methods are restricted by limited types of tissues that can be cleared with each individual protocol, which inevitably led to the presence of blind-spots within whole body or body parts imaging. Hard tissues including bones and teeth are still the most difficult organs to be cleared. In addition, loss of endogenous fluorescence remains a major concern for solvent-based clearing methods. Here, we developed a polyethylene glycol (PEG)-associated solvent system (PEGASOS), which rendered nearly all types of tissues transparent and preserved endogenous fluorescence. Bones and teeth could be turned nearly invisible after clearing. The PEGASOS method turned the whole adult mouse body transparent and we were able to image an adult mouse head composed of bones, teeth, brain, muscles, and other tissues with no blind areas. Hard tissue transparency enabled us to reconstruct intact mandible, teeth, femur, or knee joint in 3-D. In addition, we managed to image intact mouse brain at sub-cellular resolution and to trace individual neurons and axons over a long distance. We also visualized dorsal root ganglions directly through vertebrae. Finally, we revealed the distribution pattern of neural network in 3-D within the marrow space of long bone. These results suggest that the PEGASOS method is a useful tool for general biomedical research.

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製品内容

Sigma-Aldrich
水酸化ナトリウム, BioXtra, ≥98% (acidimetric), pellets (anhydrous)
Sigma-Aldrich
抗ラミニン ウサギ宿主抗体, 0.5 mg/mL, affinity isolated antibody, buffered aqueous solution
Millipore
エチレンジアミン四酢酸 二ナトリウム塩 二水和物
Sigma-Aldrich
N,N,N′,N′-テトラキス(2-ヒドロキシプロピル)エチレンジアミン, 98%
Sigma-Aldrich
抗アクチンα-平滑筋-FITC抗体、マウスモノクローナル マウス宿主抗体, clone 1A4, purified from hybridoma cell culture
Sigma-Aldrich
安息香酸ベンジル, ReagentPlus®, ≥99.0%