title: Coordinate transformation based cryo-correlative methods for electron tomography and focused ion beam milling.
abstract: Correlative microscopy allows imaging of the same feature over multiple length scales, combining light microscopy with high resolution information provided by electron microscopy. We demonstrate two procedures for coordinate transformation based correlative microscopy of vitrified biological samples applicable to different imaging modes. The first procedure aims at navigating cryo-electron tomography to cellular regions identified by fluorescent labels. The second procedure, allowing navigation of focused ion beam milling to fluorescently labeled molecules, is based on the introduction of an intermediate scanning electron microscopy imaging step to overcome the large difference between cryo-light microscopy and focused ion beam imaging modes. These methods make it possible to image fluorescently labeled macromolecular complexes in their natural environments by cryo-electron tomography, while minimizing exposure to the electron beam during the search for features of interest.
authors: Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors: Yoshiyuki
authors: Y
authors: Fukuda
authors: Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors: Nikolas
authors: N
authors: Schrod
authors: Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors: Miroslava
authors: M
authors: Schaffer
authors: Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors: Li Rebekah
authors: LR
authors: Feng
authors: Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors: Wolfgang
authors: W
authors: Baumeister
authors: Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany. Electronic address: vladan@biochem.mpg.de.
authors: Vladan
authors: V
authors: Lucic
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods*; D055032:Electron Microscope Tomography / Q000379:methods*; D004583:Electrons; D005456:Fluorescent Dyes / Q000737:chemistry; D046911:Macromolecular Substances / Q000737:chemistry; D008856:Microscopy, Fluorescence / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D046911:Macromolecular Substances
keywords: Correlative microscopy; Cryo-electron tomography; Electron tomography; Focused ion beam