title: Cellular nanoimaging by cryo electron tomography.
abstract: Cryo electron tomography is a technique that allows visualization of biological specimens in three dimensions with nanometer resolution. For cryo immobilized life sciences samples it can reveal cellular morphology, the shape of membranous structures, and depict internal macromolecular arrangements and large proteins. Cryo electron tomography is a unique technique in structural biology research because it is the only tool that enables direct visualization of the cellular space at molecular resolution. Here we present the methods that we apply in our lab to perform cellular cryo electron tomography, which require expertise on cell biology for cell growth, physics for electron microscopy, and image processing for reconstruction and 3D visualization. We define the instrumentation, materials, and protocols for cryo electron tomography of whole cells, including cell growth, specimen vitrification, microscope alignments, data acquisition, tomographic image reconstruction, and 3D visualization techniques.
authors.affiliation: Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands. r.i.koning@lumc.nl
authors.forename: Roman I
authors.initials: RI
authors.lastname: Koning
authors.forename: Abraham J
authors.initials: AJ
authors.lastname: Koster
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D020285:Cryoelectron Microscopy / Q000295:instrumentation / Q000379:methods*; D055032:Electron Microscope Tomography / Q000379:methods*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D051379:Mice; D057054:Molecular Imaging / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D011309:Preservation, Biological; D012984:Software; D013223:Statistics as Topic; D058989:Vitrification