Computational And Experimental Techniques For Expanding The Applicability Of Cryo Electron Tomography To Cell Biology
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Computational And Experimental Techniques For Expanding The Applicability Of Cryo Electron Tomography To Cell Biology
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Author : Matthew Croxford
language : en
Publisher:
Release Date : 2022
Computational And Experimental Techniques For Expanding The Applicability Of Cryo Electron Tomography To Cell Biology written by Matthew Croxford and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2022 with categories.
The field of cell biology originated with, and has always been inextricably tied to, the microscope. Direct visualization has always been a key technique for understanding cell organization and behavior. Today, cell biologists have microscopes many orders of magnitude more powerful than their progenitors, enabling the direct visualization of cells and their constituent parts down to sub-nanometer resolution. Cryo-focused ion beam milling combined with cryo-elecyron tomography (cryo-FIB-ET) is a technique in which cells are rapidly frozen to preserve them in their native state, followed by focused ion beam (FIB) thinning of the frozen cells, which allows for the collection of high resolution transmission electron microscopy data. When these electron micrographs are collected over a range of different tilt angles, a full three-dimensional reconstruction of the cellular interior can be made and analyzed. However, despite the unprecedented level of detail available to cell biologists through cryo-FIB-ET, several challenges hinder its applicability to cell biology. On the experimental side, commonly used cryo-FIB-ET compatible substrates were not designed with cell biology in mind, and have features that make them sub-optimal for this purpose. Furthermore, limitations in the range of tilts at which data may be collected lead to missing information in the three-dimensional reconstructions that cause distortions and make the resulting volumes more difficult to interpret. The work presented in this dissertation focuses on both of these issues in an attempt to increase the utility of cryo-FIB-ET for cell biologists and expand the range of questions that may be addressed with the technique.
Correlative Imaging
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Author : Paul Verkade
language : en
Publisher: John Wiley & Sons
Release Date : 2019-11-04
Correlative Imaging written by Paul Verkade and has been published by John Wiley & Sons this book supported file pdf, txt, epub, kindle and other format this book has been release on 2019-11-04 with Science categories.
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.
Single Particle Cryo Electron Microscopy
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Author : Joachim Frank
language : en
Publisher: World Scientific Publishing Company
Release Date : 2017-12-31
Single Particle Cryo Electron Microscopy written by Joachim Frank and has been published by World Scientific Publishing Company this book supported file pdf, txt, epub, kindle and other format this book has been release on 2017-12-31 with Electron microscopy categories.
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.
The Eukaryotic Ribosome
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Author : Heinz Bielka
language : en
Publisher: Walter de Gruyter GmbH & Co KG
Release Date : 1982-12-31
The Eukaryotic Ribosome written by Heinz Bielka and has been published by Walter de Gruyter GmbH & Co KG this book supported file pdf, txt, epub, kindle and other format this book has been release on 1982-12-31 with Science categories.
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The Resolution Revolution Recent Advances In Cryoem
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Author :
language : en
Publisher: Academic Press
Release Date : 2016-08-26
The Resolution Revolution Recent Advances In Cryoem written by and has been published by Academic Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2016-08-26 with Science categories.
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
Liquid Cell Electron Microscopy
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Author : Frances M. Ross
language : en
Publisher: Cambridge University Press
Release Date : 2017
Liquid Cell Electron Microscopy written by Frances M. Ross and has been published by Cambridge University Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2017 with Science categories.
2.6.2 Electrodes for Electrochemistry
Biomedical Electron Microscopy
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Author : Arvid B. Maunsbach
language : en
Publisher: Elsevier
Release Date : 1998-11-03
Biomedical Electron Microscopy written by Arvid B. Maunsbach and has been published by Elsevier this book supported file pdf, txt, epub, kindle and other format this book has been release on 1998-11-03 with Science categories.
This comprehensive reference illustrates optimal preparation methods in biological electron microscopy compared with common methodological problems. Not only will the basic methodologies of transmission electron microscopy like fixation, microtomy, and microscopy be presented, but the authors also endeavor to illustrate more specialized techniques such as negative staining, autoradiography, cytochemistry, immunoelectron microscopy, and computer-assisted image analysis. Authored by the key leaders in the biological electron microscopy field Illustrates both optimal and suboptimal or artifactual results in a variety of electron microscopy disciplines Introduces students on how to read and interpret electron micrographs
Visualizing Chemistry
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Author : National Research Council
language : en
Publisher: National Academies Press
Release Date : 2006-06-01
Visualizing Chemistry written by National Research Council and has been published by National Academies Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2006-06-01 with Science categories.
Scientists and engineers have long relied on the power of imaging techniques to help see objects invisible to the naked eye, and thus, to advance scientific knowledge. These experts are constantly pushing the limits of technology in pursuit of chemical imagingâ€"the ability to visualize molecular structures and chemical composition in time and space as actual events unfoldâ€"from the smallest dimension of a biological system to the widest expanse of a distant galaxy. Chemical imaging has a variety of applications for almost every facet of our daily lives, ranging from medical diagnosis and treatment to the study and design of material properties in new products. In addition to highlighting advances in chemical imaging that could have the greatest impact on critical problems in science and technology, Visualizing Chemistry reviews the current state of chemical imaging technology, identifies promising future developments and their applications, and suggests a research and educational agenda to enable breakthrough improvements.
Electron Crystallography Of Biological Macromolecules
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Author : Robert M. Glaeser
language : en
Publisher: Oxford University Press, USA
Release Date : 2007
Electron Crystallography Of Biological Macromolecules written by Robert M. Glaeser and has been published by Oxford University Press, USA this book supported file pdf, txt, epub, kindle and other format this book has been release on 2007 with Science categories.
This book provides a complete introduction to all major topics needed in order to use electron microscopy as a research tool in structural biology. Written by a group of 5 well-known pioneers of the field of electron cryo-microscopy of biological macromolecules, this book offers a depth of knowledge and expertise that could only be replicated from the primary literature with great difficulty.
Cryotechniques In Biological Electron Microscopy
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Author : Rudolf A. Steinbrecht
language : en
Publisher: Springer Science & Business Media
Release Date : 2012-12-06
Cryotechniques In Biological Electron Microscopy written by Rudolf A. Steinbrecht and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012-12-06 with Science categories.
To preserve tissue by freezing is an ancient concept going back pre sumably to the practice of ice-age hunters. At first glance, it seems as simple as it is attractive: the dynamics of life are frozen in, nothing is added and nothing withdrawn except thermal energy. Thus, the result should be more life-like than after poisoning, tan ning and drying a living cell as we may rudely call the conventional preparation of specimens for electron microscopy. Countless mishaps, however, have taught electron microscopists that cryotechniques too are neither simple nor necessarily more life-like in their outcome. Not too long ago, experts in cryotechniques strictly denied that a cell could truly be vitrified, i.e. that all the solutes and macro molecules could be fixed within non-crystalline, glass-like solid water without the dramatic shifts and segregation effects caused by crystallization. We now know that vitrification is indeed pos sible. Growing insight into the fundamentals of the physics of water and ice, as well as increasing experience of how to cool cells rapidly enough have enlivened the interest in cryofixation and pro duced a wealth of successful applications.