Visualization of macromolecular complexes using cryo-electron microscopy with FEI Tecnai transmission electron microscopes
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作者:
Grassucci, Robert A.
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Columbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USAColumbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA
Grassucci, Robert A.
[1
]
Taylor, Derek
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Wadsworth Ctr, Hlth Res Inc, Albany, NY 12201 USAColumbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA
Taylor, Derek
[2
]
Frank, Joachim
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Columbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA
Columbia Univ, Dept Biol Sci, New York, NY 10027 USAColumbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA
Frank, Joachim
[1
,3
]
机构:
[1] Columbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA
[2] Wadsworth Ctr, Hlth Res Inc, Albany, NY 12201 USA
[3] Columbia Univ, Dept Biol Sci, New York, NY 10027 USA
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).
机构:
Columbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USAColumbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA
Grassucci, Robert A.
;
Taylor, Derek J.
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机构:
Wadsworth Ctr, Albany, NY 12201 USAColumbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA
Taylor, Derek J.
;
Frank, Joachim
论文数: 0引用数: 0
h-index: 0
机构:
Columbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA
Wadsworth Ctr, Albany, NY 12201 USA
Columbia Univ, Dept Biol Sci, New York, NY 10027 USAColumbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA
机构:
Columbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USAColumbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA
Grassucci, Robert A.
;
Taylor, Derek J.
论文数: 0引用数: 0
h-index: 0
机构:
Wadsworth Ctr, Albany, NY 12201 USAColumbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA
Taylor, Derek J.
;
Frank, Joachim
论文数: 0引用数: 0
h-index: 0
机构:
Columbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA
Wadsworth Ctr, Albany, NY 12201 USA
Columbia Univ, Dept Biol Sci, New York, NY 10027 USAColumbia Univ Coll Phys & Surg, Howard Hughes Med Inst, Dept Biochem & Mol Biophys, New York, NY 10032 USA