Challenges and Solutions in Sample Preparation for High-Resolution Cryo-Electron Microscopy PDF Download

Author: Felix Weis
Publisher: Frontiers Media SA
ISBN: 283250860X
Category : Science
Languages : en
Pages : 120

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Author: Peter Anthony Leong
Publisher:
ISBN:
Category : Electronic dissertations
Languages : en
Pages : 214

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Author:
Publisher: Academic Press
ISBN: 0080956955
Category : Science
Languages : en
Pages : 466

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Book Description
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

Author:
Publisher:
ISBN:
Category : Cryobiology
Languages : en
Pages : 0

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Book Description
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.

Author:
Publisher: Academic Press
ISBN: 0128054352
Category : Science
Languages : en
Pages : 488

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Book Description
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

Author: Joachim Frank
Publisher: World Scientific Publishing Company
ISBN: 9789813234857
Category : Electron microscopy
Languages : en
Pages : 0

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Book Description
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.

Author: Jeanne Ayache
Publisher: Springer Science & Business Media
ISBN: 9781441959744
Category : Technology & Engineering
Languages : en
Pages : 338

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Book Description
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.

Author: Paul Verkade
Publisher: John Wiley & Sons
ISBN: 1119086450
Category : Science
Languages : en
Pages : 245

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Book Description
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.

Author: Xueting Zhou
Publisher:
ISBN:
Category :
Languages : en
Pages : 47

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Book Description
With the development of equipment and software, cryo electronic microscopy (cryoEM) was wildly used in structural biology to investigate biomolecular structure at atomic resolution. However, the sample quality is a barrier to pursuing higher resolution, including uneven distribution problems, heterogonous problems, and preferred orientation problems. Electron cryotomography (cryoET), a special application of cryoEM, have the advantage to resolve the 3D structure with a series of 2D tilting image at the same location, which is good to study the sample distribution in the thin ice layer. In this thesis, sample distribution was visualized by cryoET, and surfactant fluorinated fos choline 8 was used to manipulate the sample distribution. Besides, the principle underlying these problems were addressed based on calculation. Surface tension was the primary explanation for these problems.

Author: Annie Cavalier
Publisher: CRC Press
ISBN: 9780849372278
Category : Science
Languages : en
Pages : 0

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Book Description
While new discoveries have led to much dramatic growth in cryo-electron microscopy, researchers will never be able to take full advantage if they lack access to the details that make these techniques understandable and applicable. The Handbook of Cryo-Preparation Methods for Electron Microscopy provides researchers with a complete reference that will show them how to equip their labs with the right materials and methods to take full advantage of the latest advances. This pioneering work brings together a group of internationally renowned researchers, some the very inventors of the methods they describe, to share their knowledge and recipes. Taking care to explain the history behind the techniques and to demonstrate their use, this book presents the latest theory, principles, and protocols supplemented by hundreds of illustrations. Contributions to this handbook – Ö Describe cryo-methods aimed at perfect preservation for fine structural analysis Ö Teach how to arrest physiological processes by cryo-fixation Ö Reveal the secrets for high-resolution snapshots of life by CEMOVIS Ö Cover applications of cryo-methods such as electron crystallography, 2D/3D structure analysis, protein localization, and cryo-electron tomography Ö Demonstrate the use of vitreous water as an intermediate step for localization of biological ions and molecules Ö Present hybrid methods of freeze-substitution and freeze-drying for immunolabeling and determining molecular geography Ö Illustrate freeze-fracture, cryo-ultramicrotomy, and resin embedding Ö Highlight the Tokuyasu method and the new rehydration technique for highly efficient immunolabeling Ö Include summary tables to compare and obtain appropriate criteria Includes a wealth of immediate reference material for daily use With clear, step-by-step recipes and lists of tools, ingredients, and suppliers, this handbookprovides researchers with the knowledge and techniques to adopt the latest cryo-methods to their current research.