Single-particle cryo-EM of biological macromolecules /

This edited book is written for students, postdocs and established investigators who want to enter the field of single-particle cryo-EM. This is a recently developed method to determine high-resolution structures of biological macromolecules. A major strength is the fact that cryo-EM does not requir...

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Detalles Bibliográficos
Clasificación:Libro Electrónico
Autores principales: Glaeser, Robert M. (Autor), Nogales, Eva (Autor), Chiu, Wah (Autor)
Formato: Electrónico eBook
Idioma:Inglés
Publicado: Bristol [England] (Temple Circus, Temple Way, Bristol BS1 6HG, UK) : IOP Publishing, [2021]
Colección:IOP (Series). Release 21.
Biophysical Society-IOP series.
IOP ebooks. 2021 collection.
Temas:
Macromolecules > Analysis.
Biomolecules > Analysis.
High resolution imaging.
Biophysics.
SCIENCE / Life Sciences / Biophysics.
Acceso en línea:Texto completo
Tabla de Contenidos:
  • 1. Introduction and overview
  • 1.1. Visualizing biological molecules to understand life's principles
  • 1.2. Recovery of 3D structures from images of weak-phase objects
  • 2. Sample preparation
  • 2.1. Overview
  • 2.2. Initial screening of samples in negative stain
  • 2.3. Standard method of making grids for cryo-EM
  • 2.4. Requirement to make very thin specimens for cryo-EM
  • 2.5. Current strategies for optimizing preparation of cryo-grids
  • 3. Data collection
  • 3.1. Overview
  • 3.2. Radiation damage in cryo-EM
  • 3.3. Low-dose protocols for recording images
  • 3.4. Practical considerations : defocus, stigmation, coma-free illumination, and phase plates
  • 3.5. Practical considerations : movie-mode data acquisition
  • 4. Data processing
  • 4.1. Overview
  • 4.2. Automated extraction of particles
  • 4.3. CTF estimation and image correction (restoration)
  • 4.4. Merging data from structurally homogeneous subsets
  • 4.5. 3D classification of structurally heterogeneous particles
  • 4.6. Preferred orientation : how to recognize and deal with adverse effects
  • 4.7. B factors and map sharpening
  • 4.8. Optical aberrations and Ewald sphere curvature
  • 5. Map validation
  • 5.1. Overview
  • 5.2. Measures of resolution : FSC and local resolution
  • 5.3. Recognizing the effect of bias and over-fitting
  • 5.4. Estimates of alignment accuracy
  • 5.5. Discussion
  • 6. Model building and validation
  • 6.1. Overview
  • 6.2. Using known components or homologs : model building
  • 6.3. Building atomistic models in cryo-EM density maps
  • 6.4. Quality evaluation of cryo-EM map-derived models
  • 6.5. How algorithms from crystallography are helping electron cryo-microscopy
  • 6.6. Archiving structures and data.

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