title: Hybrid fluorescence and electron cryo-microscopy for simultaneous electron and photon imaging.
abstract: Integration of fluorescence light and transmission electron microscopy into the same device would represent an important advance in correlative microscopy, which traditionally involves two separate microscopes for imaging. To achieve such integration, the primary technical challenge that must be solved regards how to arrange two objective lenses used for light and electron microscopy in such a manner that they can properly focus on a single specimen. To address this issue, both lateral displacement of the specimen between two lenses and specimen rotation have been proposed. Such movement of the specimen allows sequential collection of two kinds of microscopic images of a single target, but prevents simultaneous imaging. This shortcoming has been made up by using a simple optical device, a reflection mirror. Here, we present an approach toward the versatile integration of fluorescence and electron microscopy for simultaneous imaging. The potential of simultaneous hybrid microscopy was demonstrated by fluorescence and electron sequential imaging of a fluorescent protein expressed in cells and cathodoluminescence imaging of fluorescent beads.
authors.affiliation: JEOL Ltd, Musashino, Akishima, Tokyo, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Iijima
authors.affiliation: National Institute for Physiological Sciences, National Institutes of Natural Sciences, Higashiyama, Myodaiji, Okazaki, Japan.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Fukuda
authors.affiliation: Terabase Inc. Higashiyama, Myodaiji, Okazaki, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Arai
authors.affiliation: Hamamatsu University School of Medicine, Handayama, Hamamatsu, Japan.
authors.forename: Susumu
authors.initials: S
authors.lastname: Terakawa
authors.affiliation: Tokyo Institute of Technology, Ookayama, Tokyo, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Yamamoto
authors.affiliation: National Institute for Physiological Sciences, National Institutes of Natural Sciences, Higashiyama, Myodaiji, Okazaki, Japan. Electronic address: nagayama@nips.ac.jp.
authors.forename: Kuniaki
authors.initials: K
authors.lastname: Nagayama
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D020285:Cryoelectron Microscopy / Q000295:instrumentation* / Q000379:methods*; D004583:Electrons; D005453:Fluorescence; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods*; D017785:Photons; D048328:Potoroidae
keywords: Cathodoluminescence; Correlative microscopy; Cryo-microscopy; Fluorescence microscope; Fluorescent protein; Transmission electron microscope