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Observation of Bacteriophage Ultrastructure by Cryo-Electron Microscopy

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Bacteriophage Therapy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2734))

Abstract

Transmission electron microscopy (TEM) is an ideal method to observe and determine the structure of bacteriophages. From early studies by negative staining to the present atomic structure models derived from cryo-TEM, bacteriophage detection, classification, and structure determination have been mostly done by electron microscopy. Although embedding in metal salts has been a routine method for virus observation for many years, the preservation of bacteriophages in a thin layer of fast frozen buffer has proven to be the most convenient preparation method for obtaining images using cryo-electron microscopy (cryo-EM). In this technique, frozen samples are observed at liquid nitrogen temperature, and the images are acquired using different recording media. The incorporation of direct electron detectors has been a fundamental step in achieving atomic resolution images of a number of viruses. These projection images can be numerically combined using different approaches to render a three-dimensional model of the virus. For those viral components exhibiting any symmetry, averaging can nowadays achieve atomic structures in most cases. Image processing methods have also evolved to improve the resolution in asymmetric viral components or regions showing different types of symmetries (symmetry mismatch).

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Authors and Affiliations

  1. Department of Structure of Macromolecules, Centro Nacional de Biotecnología, CSIC, Madrid, Spain

    Ana Cuervo, Patricia Losana & José L. Carrascosa

Authors
  1. Ana Cuervo
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  2. Patricia Losana
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  3. José L. Carrascosa
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Corresponding author

Correspondence to Ana Cuervo .

Editor information

Editors and Affiliations

  1. Centre of Biological Engineering, University of Minho, Braga, Portugal

    Joana Azeredo

  2. International Iberian Nanotechnology Laboratory, Braga, Portugal

    Sanna Sillankorva

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Cuervo, A., Losana, P., Carrascosa, J.L. (2024). Observation of Bacteriophage Ultrastructure by Cryo-Electron Microscopy. In: Azeredo, J., Sillankorva, S. (eds) Bacteriophage Therapy. Methods in Molecular Biology, vol 2734. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3523-0_2

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  • DOI: https://doi.org/10.1007/978-1-0716-3523-0_2

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3522-3

  • Online ISBN: 978-1-0716-3523-0

  • eBook Packages: Springer Protocols

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