title: Visualization of macromolecular complexes using cryo-electron microscopy with FEI Tecnai transmission electron microscopes.
abstract: This protocol details the steps used for visualizing the frozen-hydrated grids as prepared following the accompanying protocol entitled 'Preparation of macromolecular complexes for visualization using cryo-electron microscopy.' This protocol describes how to transfer the grid to the microscope using a standard cryo-transfer holder or, alternatively, using a cryo-cartridge loading system, and how to collect low-dose data using an FEI Tecnai transmission electron microscope. This protocol also summarizes and compares the various options that are available in data collection for three-dimensional (3D) single-particle reconstruction. These options include microscope settings, choice of detectors and data collection strategies both in situations where a 3D reference is available and in the absence of such a reference (random-conical and common lines).
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Howard Hughes Medical Institute, Columbia University College of Physicians and Surgeons, 630 West 168th Street, New York, New York 10032, USA.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Grassucci
authors.forename: Derek
authors.initials: D
authors.lastname: Taylor
authors.forename: Joachim
authors.initials: J
authors.lastname: Frank
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods*; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional / Q000379:methods*; D046911:Macromolecular Substances / Q000032:analysis*; D046529:Microscopy, Electron, Transmission / Q000379:methods*
chemical_list: D046911:Macromolecular Substances