Abteilung Mikrobiologie, CellNanOs-Center of Cellular Nanoanalytics Osnabrück, Fachbereich Biologie/Chemie, Universität Osnabrück, Osnabrück, Germany.
Abteilung Mikrobiologie, Fachbereich Biologie/Chemie, Universität Osnabrück, Osnabrück, Germany.
Methods Mol Biol. 2021;2182:103-115. doi: 10.1007/978-1-0716-0791-6_10.
Live cell fluorescence imaging is the method of choice to visualize dynamic cellular processes in time and space, such as adhesion to and invasion of polarized epithelial cells by Salmonella enterica sv. Typhimurium. Scanning electron microscopy provides highest resolution of surface structures of infected cells, providing ultrastructure of the apical side of host cells and infecting Salmonella. Combining both methods toward correlative light and scanning electron microscopy (CLSEM) enables new insights in adhesion and invasion mechanisms regarding dynamics over time, and high spatial resolution with precise time lines. To correlate fast live cell imaging of polarized monolayer cells with scanning electron microscopy, we developed a robust method by using gold mesh grids as convenient CLSEM carriers for standard microscopes. By this, we were able to unravel the morphology of the apical structures of monolayers of polarized epithelial cells at distinct time points during Salmonella infection.
活细胞荧光成像是观察动态细胞过程的首选方法，可实时显示细胞内的动态过程，例如沙门氏菌属血清型 Typhimurium 对极化上皮细胞的黏附和侵袭。扫描电子显微镜可提供最高分辨率的感染细胞表面结构，呈现宿主细胞顶端和感染沙门氏菌的超微结构。将这两种方法（共聚焦激光扫描和扫描电子显微镜，CLSEM）相结合，可以深入了解黏附和侵袭机制的动态变化，获得具有高空间分辨率和精确时间线的信息。为了将极化单层细胞的快速活细胞成像与扫描电子显微镜相关联，我们开发了一种稳健的方法，使用金网格作为便捷的 CLSEM 载具，用于标准显微镜。通过这种方法，我们能够在沙门氏菌感染的不同时间点揭示极化上皮细胞单层的顶端结构的形态。
