Structural Study of Heterogeneous Biological Samples by Cryoelectron Microscopy and Image Processing
Structural Study of Heterogeneous Biological Samples by Cryoelectron Microscopy and Image Processing

Author: H.E. White

Publisher: Createspace Independent Publishing Platform

Published: 2017-04-21

Total Pages: 30

ISBN-13: 9781545497548

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In living organisms, biological macromolecules are intrinsically flexible and naturally exist in multiple conformations. Modern electron microscopy, especially at liquid nitrogen temperatures (cryo-EM), is able to visualise biocomplexes in nearly native conditions and in multiple conformational states. The advances made during the last decade in electronic technology and software development have led to the revelation of structural variations in complexes and also improved the resolution of EM structures. Nowadays, structural studies based on single particle analysis (SPA) suggests several approaches for the separation of different conformational states and therefore disclosure of the mechanisms for functioning of complexes. The task of resolving different states requires the examination of large datasets, sophisticated programs, and significant computing power. Some methods are based on analysis of two-dimensional images, while others are based on three-dimensional studies. In this review, we describe the basic principles implemented in the various techniques that are currently used in the analysis of structural conformations and provide some examples of successful applications of these methods in structural studies of biologically significant complexes.

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.

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.

Biomolecular and Bioanalytical Techniques
Biomolecular and Bioanalytical Techniques

Author: Vasudevan Ramesh

Publisher: John Wiley & Sons

Published: 2019-06-10

Total Pages: 576

ISBN-13: 1119483964

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An essential guide to biomolecular and bioanalytical techniques and their applications Biomolecular and Bioanalytical Techniques offers an introduction to, and a basic understanding of, a wide range of biophysical techniques. The text takes an interdisciplinary approach with contributions from a panel of distinguished experts. With a focus on research, the text comprehensively covers a broad selection of topics drawn from contemporary research in the fields of chemistry and biology. Each of the internationally reputed authors has contributed a single chapter on a specific technique. The chapters cover the specific technique’s background, theory, principles, technique, methodology, protocol and applications. The text explores the use of a variety of analytical tools to characterise biological samples. The contributors explain how to identify and quantify biochemically important molecules, including small molecules as well as biological macromolecules such as enzymes, antibodies, proteins, peptides and nucleic acids. This book is filled with essential knowledge and explores the skills needed to carry out the research and development roles in academic and industrial laboratories. A technique-focused book that bridges the gap between an introductory text and a book on advanced research methods Provides the necessary background and skills needed to advance the research methods Features a structured approach within each chapter Demonstrates an interdisciplinary approach that serves to develop independent thinking Written for students in chemistry, biological, medical, pharmaceutical, forensic and biophysical sciences, Biomolecular and Bioanalytical Techniques is an in-depth review of the most current biomolecular and bioanalytical techniques in the field.

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

Cryo-EM Part B: 3-D Reconstruction
Cryo-EM Part B: 3-D Reconstruction

Author:

Publisher: Academic Press

Published: 2010-09-30

Total Pages: 472

ISBN-13: 0123849926

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This volume 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

cryoEM
cryoEM

Author: Tamir Gonen

Publisher: Humana

Published: 2020-12-27

Total Pages: 350

ISBN-13: 9781071609651

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This volume details the most up-to-date cryo-EM techniques from leading researchers. Chapters are organized into four parts with emphasis on electron cryotomography, single particle analysis, and the crystal based cryo-EM methods of 2D electron crystallography, and MicroED for the study of 3D crystals. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, CryoEM: Methods and Protocols aims to serve as an excellent resource on cryo-EM and can serve as the foundation for new researchers to this growing field in structural biology.

Structural Virology
Structural Virology

Author: Mavis Agbandje-McKenna

Publisher: Royal Society of Chemistry

Published: 2010-11-11

Total Pages: 389

ISBN-13: 184973223X

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Over the last ten years, much effort has been devoted to improving the biophysical techniques used in the study of viruses. This has resulted in the visualization of these large macromolecular assemblages at atomic level, thus providing the platform for functional interpretation and therapeutic design. Structural Virology covers a wide range of topics and is split into three sections. The first discusses the vast biophysical methodologies used in structural virology, including sample production and purification, confocal microscopy, mass spectrometry, negative-stain and cryo-electron microscopy, X-ray crystallography and nuclear magnetic resonance spectroscopy. The second discusses the role of virus capsid protein structures in determining the functional roles required for receptor recognition, cellular entry, capsid assembly, genome packaging and mechanisms of host immune system evasion. The last section discusses therapeutic strategies based on virus protein structures, including the design of antiviral drugs and the development of viral capsids as vehicles for foreign gene delivery. Each topic covered will begin with a review of the current literature followed by a more detailed discussion of experimental procedures, a step in the viral life cycle, or strategies for therapeutic development. With contributions from experts in the field of structural biology and virology this exceptional monograph will appeal to biomedical scientists involved in basic and /or applied research on viruses. It also provides up-to-date reference material for students entering the field of structural virology as well as scientists already familiar with the area.

Electron Microscopy
Electron Microscopy

Author: Masashi Arita

Publisher: BoD – Books on Demand

Published: 2019-10-02

Total Pages: 102

ISBN-13: 1838818820

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TEM and SEM have contributed greatly to the progress of various research fields, which has been accelerated in the last few decades by highly functional electron microscopes and microscopy. In this tide of microscopy, various microscopic methods have been developed to make clear many unsolved problems, e.g. pulse beam TEM, environmental microscopy, correlative microscopy, etc. In this book, a number of reviews have been collected concerning these subjects. We think that the content in each chapter is impressive, and we hope this book will contribute to future advances in electron microscopy, materials science, and biomedicine.