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    Nat Protoc. 2008;3(2):330-9.

    Visualization of macromolecular complexes using cryo-electron microscopy with FEI Tecnai transmission electron microscopes.

    Grassucci RA, Taylor D, Frank J.

    Source

    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.

    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).

    PMID:
    18274535
    [PubMed - indexed for MEDLINE]
    PMCID:
    PMC2654234
    Free PMC Article

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