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Expansion microscopy for the analysis of centrioles and cilia
Journal of Microscopy ( IF 1.5 ) Pub Date : 2019-11-19 , DOI: 10.1111/jmi.12841
N Sahabandu 1 , D Kong 1 , V Magidson 2 , R Nanjundappa 3 , C Sullenberger 1 , M R Mahjoub 3 , J Loncarek 1
Affiliation  

Centrioles are vital cellular structures that organise centrosomes and cilia. Due to their subresolutional size, centriole ultrastructural features have been traditionally analysed by electron microscopy. Here we present an adaptation of magnified analysis of the proteome expansion microscopy method, to be used for a robust analysis of centriole number, duplication status, length, structural abnormalities and ciliation by conventional optical microscopes. The method allows the analysis of centriole's structural features from large populations of adherent and nonadherent cells and multiciliated cultures. We validate the method using EM and superresolution microscopy and show that it can be used as an affordable and reliable alternative to electron microscopy in the analysis of centrioles and cilia in various cell cultures.

中文翻译:


用于分析中心粒和纤毛的膨胀显微镜



中心粒是组织中心体和纤毛的重要细胞结构。由于其亚分辨率尺寸,中心粒超微结构特征传统上通过电子显微镜进行分析。在这里,我们提出了蛋白质组扩展显微镜方法的放大分析,用于通过传统光学显微镜对中心粒数量、重复状态、长度、结构异常和纤毛进行稳健分析。该方法可以分析大量贴壁和非贴壁细胞以及多纤毛培养物的中心粒的结构特征。我们使用电子显微镜和超分辨率显微镜验证了该方法,并表明它可以作为电子显微镜的一种经济且可靠的替代品来分析各种细胞培养物中的中心粒和纤毛。
更新日期:2019-11-19
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