López Claudia S, Bouchet-Marquis Cedric, Arthur Christopher P, Riesterer Jessica L, Heiss Gregor, Thibault Guillaume, Pullan Lee, Kwon Sunjong, Gray Joe W
Oregon Health and Sciences University, Portland, OR, United States.
Thermo Fisher Scientific, Hillsboro, OR, United States.
Methods Cell Biol. 2017;140:149-164. doi: 10.1016/bs.mcb.2017.03.008. Epub 2017 Apr 19.
While fluorescence microscopy provides tools for highly specific labeling and sensitive detection, its resolution limit and lack of general contrast has hindered studies of cellular structure and protein localization. Recent advances in correlative light and electron microscopy (CLEM), including the fully integrated CLEM workflow instrument, the FEI CorrSight with MAPS, have allowed for a more reliable, reproducible, and quicker approach to correlate three-dimensional time-lapse confocal fluorescence data, with three-dimensional focused ion beam-scanning electron microscopy data. Here we demonstrate the entire integrated CLEM workflow using fluorescently tagged MCF7 breast cancer cells.
虽然荧光显微镜提供了用于高度特异性标记和灵敏检测的工具，但其分辨率限制和缺乏通用对比度阻碍了对细胞结构和蛋白质定位的研究。相关光电子显微镜（CLEM）的最新进展，包括完全集成的CLEM工作流程仪器——配备MAPS的FEI CorrSight，使得一种更可靠、可重复且更快的方法成为可能，能够将三维延时共聚焦荧光数据与三维聚焦离子束扫描电子显微镜数据相关联。在这里，我们使用荧光标记的MCF7乳腺癌细胞展示了整个集成的CLEM工作流程。
