Klena Nikolai, Maltinti Giovanni, Batman Umut, Pigino Gaia, Guichard Paul, Hamel Virginie
Human Technopole, Milan, Italy.
Department of Molecular and Cellular Biology, Section of Biology, University of Geneva, Geneva, Switzerland.
Bio Protoc. 2023 Sep 5;13(17):e4792. doi: 10.21769/BioProtoc.4792.
Expansion microscopy is an innovative method that enables super-resolution imaging of biological materials using a simple confocal microscope. The principle of this method relies on the physical isotropic expansion of a biological specimen cross-linked to a swellable polymer, stained with antibodies, and imaged. Since its first development, several improved versions of expansion microscopy and adaptations for different types of samples have been produced. Here, we show the application of ultrastructure expansion microscopy (U-ExM) to investigate the 3D organization of the green algae cellular ultrastructure, with a particular emphasis on the different types of sample fixation that can be used, as well as compatible staining procedures including membranes. Graphical overview.
扩展显微镜术是一种创新方法,它能够使用简单的共聚焦显微镜对生物材料进行超分辨率成像。该方法的原理依赖于将生物样本与可膨胀聚合物交联、用抗体染色并成像后,生物样本的物理各向同性膨胀。自首次开发以来,已经产生了几种扩展显微镜术的改进版本以及针对不同类型样本的改编方法。在这里,我们展示了超微结构扩展显微镜术(U-ExM)在研究绿藻细胞超微结构三维组织方面的应用,特别强调了可使用的不同类型样本固定方法,以及包括膜在内的兼容染色程序。图形概述。