title: Live correlative light-electron microscopy to observe molecular dynamics in high resolution.
abstract: Fluorescence microscopy (FM) is a powerful tool for observing specific molecular components in living cells, but its spatial resolution is relatively low. In contrast, electron microscopy (EM) provides high-resolution information about cellular structures, but it cannot provide temporal information in living cells. To achieve molecular selectivity in imaging at high resolution, a method combining EM imaging with live-cell fluorescence imaging, known as live correlative light-EM (CLEM), has been developed. In this method, living cells are first observed by FM, fixed in situ during the live observation and then subjected to EM observation. Various fluorescence techniques and tools can be applied for FM, resulting in the generation of various modified methods that are useful for understanding cellular structure in high resolution. Here, we review the methods of CLEM and live-cell imaging associated with CLEM (live CLEM). Such methods can greatly advance the understanding of the function of cellular structures on a molecular level, and thus are useful for medical fields as well as for basic biology.
authors.affiliation: Advanced ICT Research Institute Kobe, National Institute of Information and Communications Technology, 588-2 Iwaoka, Iwaoka-cho, Nishi-ku, Kobe 651-2492, Japan.
authors.forename: Shouhei
authors.initials: S
authors.lastname: Kobayashi
authors.affiliation: Advanced ICT Research Institute Kobe, National Institute of Information and Communications Technology, 588-2 Iwaoka, Iwaoka-cho, Nishi-ku, Kobe 651-2492, Japan.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Iwamoto
authors.affiliation: Advanced ICT Research Institute Kobe, National Institute of Information and Communications Technology, 588-2 Iwaoka, Iwaoka-cho, Nishi-ku, Kobe 651-2492, Japan Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka 560-0043, Japan tokuko@nict.go.jp.
authors.forename: Tokuko
authors.initials: T
authors.lastname: Haraguchi
mesh_terms: D045744:Cell Line, Tumor; D005456:Fluorescent Dyes; D006367:HeLa Cells; D006801:Humans; D053768:Metal Nanoparticles; D008854:Microscopy, Electron / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D056004:Molecular Dynamics Simulation; D045663:Quantum Dots; D012441:Saccharomyces cerevisiae / Q000648:ultrastructure*; D012568:Schizosaccharomyces / Q000648:ultrastructure*; D016808:Tetrahymena thermophila / Q000648:ultrastructure*
chemical_list: D005456:Fluorescent Dyes
keywords: CLEM; electron microscopy; fluorescence microscopy; live CLEM; live-cell imaging