A Primer to Single-Particle Cryo-Electron Microscopy
Cryo-electron microscopy (cryo-EM) of single-particle specimens is used to determine the structure of proteins and macromolecular complexes without the need for crystals. Recent advances in detector technology and software algorithms now allow images of unprecedented quality to be recorded and struc...
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| Published in | Cell Vol. 161; no. 3; pp. 438 - 449 |
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| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
23.04.2015
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| Subjects | |
| Online Access | Get full text |
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| Abstract | Cryo-electron microscopy (cryo-EM) of single-particle specimens is used to determine the structure of proteins and macromolecular complexes without the need for crystals. Recent advances in detector technology and software algorithms now allow images of unprecedented quality to be recorded and structures to be determined at near-atomic resolution. However, compared with X-ray crystallography, cryo-EM is a young technique with distinct challenges. This primer explains the different steps and considerations involved in structure determination by single-particle cryo-EM to provide an overview for scientists wishing to understand more about this technique and the interpretation of data obtained with it, as well as a starting guide for new practitioners. |
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| AbstractList | Cryo-electron microscopy (cryo-EM) of single-particle specimens is used to determine the structure of proteins and macromolecular complexes without the need for crystals. Recent advances in detector technology and software algorithms now allow images of unprecedented quality to be recorded and structures to be determined at near-atomic resolution. However, compared with X-ray crystallography, cryo-EM is a young technique with distinct challenges. This primer explains the different steps and considerations involved in structure determination by single-particle cryo-EM to provide an overview for scientists wishing to understand more about this technique and the interpretation of data obtained with it, as well as a starting guide for new practitioners.Cryo-electron microscopy (cryo-EM) of single-particle specimens is used to determine the structure of proteins and macromolecular complexes without the need for crystals. Recent advances in detector technology and software algorithms now allow images of unprecedented quality to be recorded and structures to be determined at near-atomic resolution. However, compared with X-ray crystallography, cryo-EM is a young technique with distinct challenges. This primer explains the different steps and considerations involved in structure determination by single-particle cryo-EM to provide an overview for scientists wishing to understand more about this technique and the interpretation of data obtained with it, as well as a starting guide for new practitioners. Cryo-electron microscopy (cryo-EM) of single-particle specimens is used to determine the structure of proteins and macromolecular complexes without the need for crystals. Recent advances in detector technology and software algorithms now allow images of unprecedented quality to be recorded and structures to be determined at near-atomic resolution. However, compared with X-ray crystallography, cryo-EM is a young technique with distinct challenges. This primer explains the different steps and considerations involved in structure determination by single-particle cryo-EM to provide an overview for scientists wishing to understand more about this technique and the interpretation of data obtained with it, as well as a starting guide for new practitioners. |
| Author | Cheng, Yifan Penczek, Pawel A. Grigorieff, Nikolaus Walz, Thomas |
| AuthorAffiliation | 3 Department of Biochemistry and Molecular Biology, The University of Texas – Houston Medical School, 6431 Fannin Street, MSB 6.220, Houston, TX 77030, USA 1 Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94158, USA 4 Department of Cell Biology and Howard Hughes Medical Institute, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA 2 Janelia Research Campus, 19700 Helix Drive, Ashburn, VA 20147, USA |
| AuthorAffiliation_xml | – name: 4 Department of Cell Biology and Howard Hughes Medical Institute, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA – name: 1 Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94158, USA – name: 2 Janelia Research Campus, 19700 Helix Drive, Ashburn, VA 20147, USA – name: 3 Department of Biochemistry and Molecular Biology, The University of Texas – Houston Medical School, 6431 Fannin Street, MSB 6.220, Houston, TX 77030, USA |
| Author_xml | – sequence: 1 givenname: Yifan surname: Cheng fullname: Cheng, Yifan – sequence: 2 givenname: Nikolaus surname: Grigorieff fullname: Grigorieff, Nikolaus – sequence: 3 givenname: Pawel A. surname: Penczek fullname: Penczek, Pawel A. – sequence: 4 givenname: Thomas surname: Walz fullname: Walz, Thomas |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/25910204$$D View this record in MEDLINE/PubMed |
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| SubjectTerms | Algorithms computer software cryo-electron microscopy Cryoelectron Microscopy - instrumentation Cryoelectron Microscopy - methods crystals Image Processing, Computer-Assisted Models, Molecular Molecular Conformation Protein Conformation proteins Proteins - chemistry Proteins - isolation & purification Proteins - ultrastructure X-ray diffraction |
| Title | A Primer to Single-Particle Cryo-Electron Microscopy |
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