利用CLARITY技术清除骨骼肌以进行光学显微镜成像。
Clearing skeletal muscle with CLARITY for light microscopy imaging.
作者信息
Milgroom Andrew, Ralston Evelyn
机构信息
Light Imaging Section, Office of Science and Technology, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, Maryland, 20892-8023, USA.
出版信息
Cell Biol Int. 2016 Apr;40(4):478-83. doi: 10.1002/cbin.10578. Epub 2016 Feb 15.
Viewing subcellular details over large tissue volumes is becoming an essential condition of the success of large-scale projects aimed at visualizing cell connections in whole organs or tissues. However, tissue opacity remains an obstacle to deep tissue imaging. This situation has brought renewed interest for techniques of tissue clearing; new protocols, such as CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging/Immunostaining/In situ hybridization-compatible Tissue-hYdrogel), have recently been developed. So far, most of the tests of these techniques have been applied to brain or other soft tissues. Here we show that CLARITY clears mouse hindlimb skeletal muscles and maintains the basic structural features of muscle and its fibers. However, tagging with fluorescent markers was not successful.
在大体积组织上观察亚细胞细节正成为旨在可视化整个器官或组织中细胞连接的大规模项目取得成功的必要条件。然而,组织不透明度仍然是深度组织成像的障碍。这种情况重新引发了人们对组织透明化技术的兴趣;最近已经开发出了新的方案,如CLARITY(清除脂质交换丙烯酰胺杂交刚性成像/免疫染色/原位杂交兼容组织水凝胶)。到目前为止,这些技术的大多数测试都应用于大脑或其他软组织。在这里我们表明,CLARITY能使小鼠后肢骨骼肌透明化,并保持肌肉及其纤维的基本结构特征。然而,用荧光标记物进行标记并不成功。