Cryo-EM Part A: Sample Preparation and Data Collection
Cryo-EM Part A: Sample Preparation and Data Collection

Author:

Publisher: Academic Press

Published: 2010-09-30

Total Pages: 466

ISBN-13: 0080956955

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Cryo-EM Part A: Sample Preparation and Data Collection is dedicated to a description of the instruments, samples, protocols, and analyses that belong to cryo-EM. It emphasizes the relatedness of the ideas, instrumentation, and methods underlying all cryo-EM approaches, which allow practitioners to easily move between them. Within each section, the articles are ordered according to the most common symmetry of the sample to which their methods are applied. Includes time-tested core methods and new innovations applicable to any researcher Methods included are useful to both established researchers and newcomers to the field Relevant background and reference information given for procedures can be used as a guide

Challenges in Biological Cryo-electron Microscopy
Challenges in Biological Cryo-electron Microscopy

Author:

Publisher:

Published: 2022

Total Pages: 0

ISBN-13:

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Developments in cryo-electron microscopy are creating new opportunities within structural biology and there is currently great interest in developing cryo-EM as a core tool for atomic level structural biology. Many structural techniques can give atomic or near atomic level information, but lack the ability to study proteins within a near-native environment, for example within a cellular compartment. Cryo-EM provides this opportunity, but despite the recent massive improvements in single particle cryo-EM, obtaining sub-2Å structural information is still a major challenge. Cryo-electron microscopy has undergone significant developments in microscope design, camera technology and data processing regimes, but there are significant challenges that remain and opportunities to explore, many of which must be tackled by the community as a whole, rather than by individual groups. For example, sample preparation is central to electron microscopy and is currently a significant bottleneck in many experiments, and there are significant problems with ensuring the integrity of the field in terms of dealing with inherently low signal-to-noise images. This volume brings together leading researchers from the UK and the international cryo-electron microscopy community to discuss current developments and new challenges in the field. In this volume the topics covered include: Sample preparation in single particle cryo-EM ; Pushing the limits in single particle cryo-EM ; Tomographic analysis, CLEM ; Map/model validation and machine learning in EM.

The Resolution Revolution: Recent Advances In cryoEM
The Resolution Revolution: Recent Advances In cryoEM

Author:

Publisher: Academic Press

Published: 2016-08-26

Total Pages: 488

ISBN-13: 0128054352

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cryoEM, a new volume in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers research methods and new developments in recording images, the creation, evaluation and validation of 3D maps from the images, model building into maps and refinement of the resulting atomic structures, and applications of essentially single particle methods to helical structures and to sub-tomogram averaging. Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers research methods that determine the structures of biological molecules, a vital step for understanding their function Contains the technical developments underpinning the advances of cryoEM and captures the exciting insights that have resulted

Single-particle Cryo-electron Microscopy
Single-particle Cryo-electron Microscopy

Author: Joachim Frank

Publisher: World Scientific Publishing Company

Published: 2017-12-31

Total Pages: 0

ISBN-13: 9789813234857

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The book reproduces 55 of more than 300 articles written by the author, representing milestones in methods development of single-particle cryo-EM as well as important results obtained by this technique in the study of biological macromolecules and their interactions. Importantly, neither symmetries nor ordered arrangements (as in two-dimensional crystals, helical assemblies, icosahedral viruses) are required. Although the biological applications are mainly in the area of ribosome structure and function, the elucidation of membrane channel structures and their activation and gating mechanisms are represented, as well. The book is introduced by a commentary that explains the original development of concepts, describes the contributions of the author's colleagues and students, and shows how challenges were overcome as the technique matured. Along the way, the ribosome served as an example for a macromolecule with intricate structure and conformational dynamics that pose challenges for three-dimensional visualization. Toward the end of the book -- bringing us to the present time -- molecular structures with near-atomic resolution are presented, and a novel type of computational analysis, manifold embedding, is introduced. Single-particle cryo-EM is currently revolutionizing structural biology, presenting a powerful alternative to X-ray crystallography as a means to solve the structure of biological macromolecules. The book presents in one place a number of articles containing key advances in mathematical and computational methods leading up to the present time. Secondly, the development of the technique over the years is reflected by ever-expanding discoveries in the field of ribosome structure and function. Thirdly, as all histories of ideas, the history of concepts pertaining to this new method of visualization is fascinating all in itself.

Sample Preparation Handbook for Transmission Electron Microscopy
Sample Preparation Handbook for Transmission Electron Microscopy

Author: Jeanne Ayache

Publisher: Springer Science & Business Media

Published: 2010-06-08

Total Pages: 338

ISBN-13: 9781441959744

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Successful transmission electron microscopy in all of its manifestations depends on the quality of the specimens examined. Biological specimen preparation protocols have usually been more rigorous and time consuming than those in the physical sciences. For this reason, there has been a wealth of scienti c literature detailing speci c preparation steps and numerous excellent books on the preparation of b- logical thin specimens. This does not mean to imply that physical science specimen preparation is trivial. For the most part, most physical science thin specimen pre- ration protocols can be executed in a matter of a few hours using straightforward steps. Over the years, there has been a steady stream of papers written on various aspects of preparing thin specimens from bulk materials. However, aside from s- eral seminal textbooks and a series of book compilations produced by the Material Research Society in the 1990s, no recent comprehensive books on thin specimen preparation have appeared until this present work, rst in French and now in English. Everyone knows that the data needed to solve a problem quickly are more imp- tant than ever. A modern TEM laboratory with supporting SEMs, light microscopes, analytical spectrometers, computers, and specimen preparation equipment is an investment of several million US dollars. Fifty years ago, electropolishing, chemical polishing, and replication methods were the principal specimen preparation me- ods.

Handbook of Sample Preparation for Scanning Electron Microscopy and X-Ray Microanalysis
Handbook of Sample Preparation for Scanning Electron Microscopy and X-Ray Microanalysis

Author: Patrick Echlin

Publisher: Springer Science & Business Media

Published: 2011-04-14

Total Pages: 329

ISBN-13: 0387857311

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Scanning electr on microscopy (SEM) and x-ray microanalysis can produce magnified images and in situ chemical information from virtually any type of specimen. The two instruments generally operate in a high vacuum and a very dry environment in order to produce the high energy beam of electrons needed for imaging and analysis. With a few notable exceptions, most specimens destined for study in the SEM are poor conductors and composed of beam sensitive light elements containing variable amounts of water. In the SEM, the imaging system depends on the specimen being sufficiently electrically conductive to ensure that the bulk of the incoming electrons go to ground. The formation of the image depends on collecting the different signals that are scattered as a consequence of the high energy beam interacting with the sample. Backscattered electrons and secondary electrons are generated within the primary beam-sample interactive volume and are the two principal signals used to form images. The backscattered electron coefficient ( ? ) increases with increasing atomic number of the specimen, whereas the secondary electron coefficient ( ? ) is relatively insensitive to atomic number. This fundamental diff- ence in the two signals can have an important effect on the way samples may need to be prepared. The analytical system depends on collecting the x-ray photons that are generated within the sample as a consequence of interaction with the same high energy beam of primary electrons used to produce images.

Correlative Imaging
Correlative Imaging

Author: Paul Verkade

Publisher: John Wiley & Sons

Published: 2019-11-04

Total Pages: 245

ISBN-13: 1119086450

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Brings a fresh point of view to the current state of correlative imaging and the future of the field This book provides contributions from international experts on correlative imaging, describing their vision of future developments in the field based on where it is today. Starting with a brief historical overview of how the field evolved, it presents the latest developments in microscopy that facilitate the correlative workflow. It also discusses the need for an ideal correlative probe, applications in proteomic and elemental analysis, interpretation methods, and how correlative imaging can incorporate force microscopy, soft x-ray tomography, and volume electron microscopy techniques. Work on placing individual molecules within cells is also featured. Correlative Imaging: Focusing on the Future offers in-depth chapters on: correlative imaging from an LM perspective; the importance of sample processing for correlative imaging; correlative light and volume EM; correlation with scanning probe microscopies; and integrated microscopy. It looks at: cryo-correlative microscopy; correlative cryo soft X-ray imaging; and array tomography. Hydrated-state correlative imaging in vacuo, correlating data from different imaging modalities, and big data in correlative imaging are also considered. Brings a fresh view to one of the hottest topics within the imaging community: the correlative imaging field Discusses current research and offers expert thoughts on the field’s future developments Presented by internationally-recognized editors and contributors with extensive experience in research and applications Of interest to scientists working in the fields of imaging, structural biology, cell biology, developmental biology, neurobiology, cancer biology, infection and immunity, biomaterials and biomedicine Part of the Wiley–Royal Microscopical Society series Correlative Imaging: Focusing on the Future will appeal to those working in the expanding field of the biosciences, correlative microscopy and related microscopic areas. It will also benefit graduate students working in microscopy, as well as anyone working in the microscopy imaging field in biomedical research.