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Cryo-electron microscopy, in combination with tomography, has emerged as a new technology for visualizing molecular structures at a resolution beyond even 1 Å. Using this technology has revealed the native molecular details of viruses, membranes, enzymes, ribosomes, and cells. This comprehensive volume brings together authoritative overviews of these methods from structural and biological perspectives. It is a must-have for researchers and graduate students, as well as those working in industry, primarily in the areas of biophysics, structural biology, crystallography, and genomics.Key Features• Focuses on the applications of cryo-EM to structural biology• Documents the importance of cryo-EM/ET approaches in studying the structural determinants of cellular organelle and membrane protein biochemistry• Reviews the applications of high-resolution structures of viruses• Emphasizes structural insights of nuclear and gene machineries• Includes a section focused entirely on the applications of cryo-EM/ET in drug discovery and therapeutic development

Correlative Light and Electron Microscopy III, Volume 140, a new volume in the Methods in Cell Biology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Topics discussed in this new release include Millisecond time-resolved CLEM, Super resolution LM und SEM of high-pressure frozen C. elegans, Preservation fluorescence, super res CLEM, APEX in Tissue, Corrsight mit IBIDI flowthrough chamber, Correlative Light Atomic Force Electronic Microscopy (CLAFEM), Atmospheric EM CLEM, and High-precision correlation, amongst other topics. Chapters in this ongoing series deal with different approaches for analyzing the same specimen using more than one imaging technique. The strengths and application area of each is presented, with this volume exploring the aspects of sample preparation of diverse biological systems for different CLEM approaches. Contains contributions from experts in the field Covered topics include targeted ultramicrotomy and high-precision correlation Presents recent advances and currently applied correlative approaches Gives detailed protocols allowing the application of workflows in one's own laboratory setting Covers CLEM approaches in the context of specific applications Aims to stimulate the use of new combinations of imaging modalities

The first book on the topic, with each chapter written by pioneers in the field, this essential resource details the fundamental theory, applications, and future developments of liquid cell electron microscopy. This book describes the techniques that have been developed to image liquids in both transmission and scanning electron microscopes, including general strategies for examining liquids, closed and open cell electron microscopy, experimental design, resolution, and electron beam effects. A wealth of practical guidance is provided, and applications are described in areas such as electrochemistry, corrosion and batteries, nanocrystal growth, biomineralization, biomaterials and biological processes, beam-induced processing, and fluid physics. The book also looks ahead to the future development of the technique, discussing technical advances that will enable higher resolution, analytical microscopy, and even holography of liquid samples. This is essential reading for researchers and practitioners alike.

This new volume of Methods in Cell Biology looks at methods for analyzing correlative light and electron microscopy (CLEM). With CLEM, people try to combine the advantages of both worlds, i.e. the dynamics information obtained by light microscopy and the ultrastructure as provided by electron microscopy. This volume contains the latest techniques on correlative microscopy showing that combining two imaging modalities provides more than each technique alone. Most importantly it includes the essential protocols, including tips, tricks and images for you to repeat these exciting techniques in your own lab. With cutting-edge material, this comprehensive collection is intended to guide researchers for years to come. - Covers sections on model systems and functional studies, imaging-based approaches and emerging studies - Chapters are written by experts in the field - Cutting-edge material - Second of two volumes dedicated to Correlative Light and Electron microscopy (CLEM)

Part of the Wiley-Royal Microscopical Society Series, this book discusses the rapidly developing cutting-edge field of low-voltage microscopy, a field that has only recently emerged due to the rapid developments in the electron optics design and image processing. It serves as a guide for current and new microscopists and materials scientists who are active in the field of nanotechnology, and presents applications in nanotechnology and research of surface-related phenomena, allowing researches to observe materials as never before.

The combination of electron microscopy with transmitted light microscopy (termed correlative light and electron microscopy; CLEM) has been employed for decades to generate molecular identification that can be visualized by a dark, electron-dense precipitate. This new volume of Methods in Cell Biology covers many areas of CLEM, including a brief history and overview on CLEM methods, imaging of intermediate stages of meiotic spindle assembly in C. elegans embryos using CLEM, and capturing endocytic segregation events with HPF-CLEM. - Covers many areas of CLEM by the best international scientists in the field - Includes a brief history and overview on CLEM methods

For the first time, fixation, resin embedding and immunocytochemistry, the subjects of innumerable publications, have been organised into a comprehensive and coherent scheme showing how the various methodologies interrelate. Commercially available resins and their mode of use are described, and the book considers the advantages for cytochemistry and immunocytochemistry of matching tissue fixation to processing and resin embedding. On-section single and double immunolabelling methods are covered, using immunocolloidal gold and immunoperoxidase.

The modem microbiologist is often a real specialist who has difficulty under­ standing and applying many of the techniques beyond those in his or her own immediate field. On the other hand, most benefits to modem microbiology are obtained when a broad spectrum of scientific approaches can be focused on a problem. In early studies, electron microscopy was pivotal in understanding bacterial and viral morphology, and we still feel that we will understand a disease better if we have seen an electron micrograph of the causative agent. Today, because there is an increased awareness of the need to understand the rela­ tionships between microbial structure and function, the electron microscope is still one of the most important tools microbiologists can use for detailed analysis of microorganisms. Often, however, the aforementioned modem microbiologist still thinks of ultrastructure as involving negative staining or ultrathin sectioning in order to get a look at the shape of a'bug.'Many of the newer ultrastructure techniques, such as gold-labeled antibody localization, freeze-fracture, X-ray microanalysis, enzyme localization, and even scanning electron microscopy, are poorly under­ stood by, and therefore forbidding to, the average microbiologist. Even many cell biologists admit to having difficulty staying in touch with current develop­ ments in the fast-moving field of electron microscopy techniques.

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

This volume, along with Part A and Part B, 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

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

In-situ high-resolution electron microscopy is a modern and powerful technique in materials research, physics, and chemistry. In-situ techniques are hardly treated in textbooks of electron microscopy. Thus, there is a need to collect the present knowledge about the techniques and achievements of in-situ electron microscopy in one book. Since high-resolution electron microscopes are available in most modern laboratories of materials science, more and more scientists or students are starting to work on this subject.In this comprehensive volume, the most important techniques and achievements of in-situ high-resolution electron microscopy will be reviewed by renowned experts. Applications in several fields of materials science will also be demonstrated.

This volume demonstrates how cellular and associated electron microscopy contributes to knowledge about biological structural information, primarily at the nanometer level. It presents how EM approaches complement both conventional structural biology (at the high end, angstrom level of resolution) and digital light microscopy (at the low end, 100-200 nanometers). - Basic techniques in transmission and scanning electron microscopy - Detailed chapters on how to use electron microscopy when dealing with specific cellular structures, such as the nucleus, cell membrane, and cytoskeleton - Discussion on electron microscopy of viruses and virus-cell interactions

Strategies for Two-Dimensional Crystallization of Proteins Using Lipid Monolayers presents an overview of different methods that lead to structure determination by electron microscopy. These methods have proven to be extremely successful, especially for elucidating the structure of membrane proteins. Electron crystallography has become an important tool for structure determination of such proteins. This book covers the different practical approaches to two-dimensional crystallization of soluble as well as membrane proteins. From there it takes the reader to equally important issues, such as sample transfer and sample preparation for electron microscopy. In addition, the text provides an introduction to membrane protein structural biology and cryo-electron crystallography, as well as an in-depth discussion on two-dimensional crystallization and surface crystallization.

Although the polymerase chain reaction has revolutionized genetic analysis by amplifying rare nucleic acid sequences, the in situ application is the only method that allows the localization of amplified signal within tissue structure. The applications of in situ polymerase chain reaction have greatly enhanced the field of investigation in many disc

This is a clear account of the application of electron-based microscopies to the study of high-Tc superconductors. Written by leading experts, this compilation provides a comprehensive review of scanning electron microscopy, transmission electron microscopy and scanning transmission electron microscopy, together with details of each technique and its applications. Introductory chapters cover the basics of high-resolution transmission electron microscopy, including a chapter devoted to specimen preparation techniques, and microanalysis by scanning transmission electron microscopy. Ensuing chapters examine identification of superconducting compounds, imaging of superconducting properties by low-temperature scanning electron microscopy, imaging of vortices by electron holography and electronic structure determination by electron energy loss spectroscopy. The use of scanning tunnelling microscopy for exploring surface morphology, growth processes and the mapping of superconducting carrier distributions is discussed. Final chapters consider applications of electron microscopy to the analysis of grain boundaries, thin films and device structures. Detailed references are included.

Hands-on experimentalists describe the cutting-edge microscopical methods needed for the effective study of plant cell biology today. These powerful techniques, all described in great detail to ensure successful experimental results, range from light microscope cytochemistry, autoradiography, and immunocytochemistry, to recent developments in fluorescence, confocal, and dark-field microscopies. Important advances in both conventional and scanning electron microscopies are also fully developed, together with such state-of-the-art ancillary techniques as high-resolution autoradiography, immunoelectron microscopy, X-ray microanalysis, and electron systems imaging. Easy-to-use and up-to-date, Methods in Plant Electron Microscopy and Cytochemistry offers today's plant scientists a first class collection of readily reproducible light and electron microscopical methods that will prove the new standard for all working in the field.

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

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.

Histological Techniques for Electron Microscopy, Second Edition, offers a practical guide for those who would study cells or tissues with an electron microscope. The book contains 11 chapters and begins with a discussion of the organization and management of an electron microscope laboratory. This is followed by separate chapters on tissue preservation; fixatives and their variations; methacrylate embedding and cross-linked plastics, microtomes and microtomy; and section mounting. Subsequent chapters deal with the techniques of'staining'for electron microscopy, photography; mounting, shadowing, and replication; and negative staining.

Fixation for Electron Microscopy presents how to better understand the effects of fixatives on the molecular structure of the cell. This book attempts to consider each aspect of fixation, including chemical interactions between fixatives and individual cellular substances. The chemistry of fixative interactions that are discussed in the book is based primarily on the reactions of a fixative with isolated proteins, lipids, nucleic acids, and carbohydrates. The book shows that the correct interpretation of information retrieved from electron micrographs depends on the knowledge of the basic principles underlying the fixation procedure. Also, the book presents the fixation of both eukaryotic and prokaryotic specimens. The special fixation conditions for plant specimens are discussed in detail and have been allotted a whole chapter. Also emphasized in this book is the connection between morphology and biochemical aspects of preparatory treatments and the chemical basis of the formation of artifacts. This topic is useful in understanding the modifications of cell structures introduced during their processing. A guide for recognizing and minimizing major artifacts and fixation faults that are usually encountered is also presented in the book. This valuable resource will prove useful to both students and professionals in the field of biology and clinical medicine. Specimen preservation researchers can also benefit from this book.

Computer Techniques for Image Processing in Electron Microscopy: Advances in Electronics and Electron Physics presents the sophisticated computer generated in processing the image. This book discusses the development of fast Fourier transform algorithms, which has led to the possibility of achieving a more reliable interpretation of electron micrographs by digital means. Organized into 10 chapters, this book begins with an overview of image formation in which the properties of the linear approximation are included. This text then reviews the available hardware and the basic mathematical methods of image processing in electron microscopy. Other chapters consider the constraints imposed on the image wave function by the objective lens aperture. This book discusses as well the properties of discrete Fourier transforms. The final chapter deals with a particular processing system called the Improc system. This book is a valuable resource for physicists and researcher workers who are interested in the study of image processing.

Basic Techniques for Transmission Electron Microscopy describes the basic techniques for transmission electron microscopy. Preparatory procedures for both eukaryotic and prokaryotic groups are presented in a step-by-step fashion, together with special preparatory methods for plant specimens and viruses. The processing of uncommon specimens and the solution of unusual, individual problems are included. This book is comprised of seven chapters and begins with a discussion on chemical fixation, with particular reference to fixatives and the hazards, precautions, and safe handling of reagents, as well as the preparation of buffers and tissue blocks. The reader is then introduced to the standard procedure for fixation, rinsing, dehydration, and embedding. Subsequent chapters focus on sectioning, cryofixation, and cryoultramicrotomy; positive and negative staining; and the use of support films. The final chapter presents a wide variety of specimens such as algae, amoeba, anthers, actin filaments, bacteria, and cells in culture. This monograph is essentially a laboratory handbook intended for students, technicians, teachers, and research scientists in biology and medicine.

Electron Microscopy of Plant Cells serves as manual or reference of major modern techniques used to prepare plant material for transmission and scanning electron microscopy. There have been other books that generally discuss electron microscope methodology. This book focuses on problem areas encountered through the presence of tough cell walls and large central vacuole. It details preparative techniques for botanical specimens. Each of the nine chapters of this book covers the basic principles, useful applications, and reliable procedures used on the method of electron microscopy. Other topics discussed in each chapter include the general preparation and straining of thin sections, quantitative morphological analysis, and enzyme cytochemistry. This book also explains the immunogold labelling, rapid-freezing methods, and ambient- and low-temperature scanning electron microscopy among others. This book will be invaluable to general scientists, biologists, botanists, and students specializing in plant anatomy.