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PEGASOS 方法对硬组织和软组织器官的组织通透化。

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

机构信息

Department of Restorative Sciences, School of Dentistry, Texas A&M University, Dallas, TX, 75246, USA.

State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, 610041, China.

出版信息

Cell Res. 2018 Aug;28(8):803-818. doi: 10.1038/s41422-018-0049-z. Epub 2018 May 29.

Abstract

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.

摘要

组织透明化技术通过将组织透明化来实现对不透明器官和组织的三维可视化。目前的组织透明化方法受到每种方法只能用于有限类型组织的限制,这不可避免地导致在整个身体或身体部位成像中存在盲点。包括骨骼和牙齿在内的硬组织仍然是最难被透明化的器官。此外,溶剂型透明化方法中内源性荧光的损失仍然是一个主要问题。在这里,我们开发了一种聚乙二醇(PEG)相关溶剂系统(PEGASOS),它使几乎所有类型的组织都变得透明,并保留了内源性荧光。经过透明化处理后,骨骼和牙齿几乎可以变得不可见。PEGASOS 方法使整个成年小鼠体透明,我们能够对由骨骼、牙齿、大脑、肌肉和其他组织组成的成年小鼠头部进行成像,没有任何盲点。硬组织的透明度使我们能够重建完整的下颌骨、牙齿、股骨或膝关节的三维结构。此外,我们成功地以亚细胞分辨率对完整的小鼠大脑进行成像,并能够追踪单个神经元和轴突的长距离。我们还可以直接通过脊椎观察背根神经节。最后,我们揭示了长骨骨髓腔内三维神经网络的分布模式。这些结果表明,PEGASOS 方法是一般生物医学研究的有用工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bff/6082844/b8a62e651b01/41422_2018_49_Fig1_HTML.jpg

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