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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 - OE Describe cryo-methods aimed at perfect preservation for fine structural analysis OE Teach how to arrest physiological processes by cryo-fixation OE Reveal the secrets for high-resolution snapshots of life by CEMOVIS OE Cover applications of cryo-methods such as electron crystallography, 2D/3D structure analysis, protein localization, and cryo-electron tomography OE Demonstrate the use of vitreous water as an intermediate step for localization of biological ions and molecules OE Present hybrid methods of freeze-substitution and freeze-drying for immunolabeling and determining molecular geography OE Illustrate freeze-fracture, cryo-ultramicrotomy, and resin embedding OE Highlight the Tokuyasu method and the new rehydration technique for highly efficient immunolabeling OE 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.
This book presents a wide variety of immuno-gold techniques for use in virus diagnosis and research. Protocols are presented for state-of-the-art techniques, including in situ hybridization, freeze substitution, and the utilization of ultra-small probes and replicas for use by virologists and electron microscopists identifying and studying viruses, their components, and replication in cells. The procedures are described by eminent scientists and are pertinent to both experienced researchers and newcomers to this field who are interested in the localization of low antigenic mass structures.
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