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Biological Specimen Preparation for Transmission Electron Microscopy

Biological Specimen Preparation for Transmission Electron Microscopy
Author: Audrey M. Glauert
Publisher: Princeton University Press
Total Pages: 349
Release: 2014-07-14
Genre: Science
ISBN: 1400865026

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This book contains all the necessary information and advice for anyone wishing to obtain electron micrographs showing the most accurate ultrastructural detail in thin sections of any type of biological specimen. The guidelines for the choice of preparative methods are based on an extensive survey of current laboratory practice. For the first time, in a textbook of this kind, the molecular events occurring during fixation and embedding are analysed in detail. The reasons for choosing particular specimen preparation methods are explained and guidance is given on how to modify established techniques to suit individual requirements. All the practical methods advocated are clearly described, with accompanying tables and the results obtainable are illustrated with many electron micrographs. Portland Press Series: Practical Methods in Electron Microscopy, Volume 17, Audrey M. Glauert, Editor Originally published in 1999. The Princeton Legacy Library uses the latest print-on-demand technology to again make available previously out-of-print books from the distinguished backlist of Princeton University Press. These editions preserve the original texts of these important books while presenting them in durable paperback and hardcover editions. The goal of the Princeton Legacy Library is to vastly increase access to the rich scholarly heritage found in the thousands of books published by Princeton University Press since its founding in 1905.


Handbook of Cryo-Preparation Methods for Electron Microscopy

Handbook of Cryo-Preparation Methods for Electron Microscopy
Author: Annie Cavalier
Publisher: CRC Press
Total Pages: 0
Release: 2008-10-02
Genre: Science
ISBN: 9780849372278

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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 – 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.


Electron Microscopy Methods and Protocols

Electron Microscopy Methods and Protocols
Author: M. A. Nasser Hajibagheri
Publisher: Springer Science & Business Media
Total Pages: 292
Release: 2008-02-02
Genre: Science
ISBN: 1592592015

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Electron Microscopy Methods and Protocols is designed for the established researcher as a manual for extending knowledge of the field. It is also for the newcomer who wishes to move into the field. A wide range of applications for the examination of cells, tissues, biological macromolecules, molecular structures, and their interactions are discussed. We have tried to gather together methods that we consider to be those most generally appli- ble to current research in both cell and molecular biology. Each chapter c- tains a set of related practical protocols with examples provided by experts who have first-hand knowledge of the techniques they describe. The individual chapters are grouped according to similarities in their specimen preparation and methodology. Methods are presented in detail, in a step-by-step fashion, using reproducible protocols the authors have personally checked. During the last decade, the scientific literature describing the use of colloidal gold as an immunocytochemical marker has increased at an ex- nential rate, and this trend is expected to continue. We have included a large number of variations on the immunogold labeling technique. In both the ne- tive staining and cryo chapters, authors emphasize the “immunological app- cations” in order to correlate as fully as possible with the emphasis on immunogold labeling in the other chapters. Electron Microscopy Methods and Protocols commences with the routine preparation of biological material for classical transmission electron microscopy involving tissue fixation, embedding, and sectioning (Chap. 1).


Handbook of Sample Preparation for Scanning Electron Microscopy and X-Ray Microanalysis

Handbook of Sample Preparation for Scanning Electron Microscopy and X-Ray Microanalysis
Author: Patrick Echlin
Publisher: Springer Science & Business Media
Total Pages: 329
Release: 2011-04-14
Genre: Technology & Engineering
ISBN: 0387857311

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Scanning electr on microscopy (SEM) and x-ray microanalysis can produce magnified images and in situ chemical information from virtually any type of specimen. The two instruments generally operate in a high vacuum and a very dry environment in order to produce the high energy beam of electrons needed for imaging and analysis. With a few notable exceptions, most specimens destined for study in the SEM are poor conductors and composed of beam sensitive light elements containing variable amounts of water. In the SEM, the imaging system depends on the specimen being sufficiently electrically conductive to ensure that the bulk of the incoming electrons go to ground. The formation of the image depends on collecting the different signals that are scattered as a consequence of the high energy beam interacting with the sample. Backscattered electrons and secondary electrons are generated within the primary beam-sample interactive volume and are the two principal signals used to form images. The backscattered electron coefficient ( ? ) increases with increasing atomic number of the specimen, whereas the secondary electron coefficient ( ? ) is relatively insensitive to atomic number. This fundamental diff- ence in the two signals can have an important effect on the way samples may need to be prepared. The analytical system depends on collecting the x-ray photons that are generated within the sample as a consequence of interaction with the same high energy beam of primary electrons used to produce images.


Electron Microscopy

Electron Microscopy
Author: John Kuo
Publisher: Humana
Total Pages: 0
Release: 2016-08-27
Genre: Medical
ISBN: 9781493960736

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This third edition of Electron Microscopy: Methods and Protocols expands upon the previous editions with current, detailed protocols on biological and molecular research techniques based on TEM and SEM as well as other closely related imaging and analytical methods. With new chapters on conventional and microwave assisted specimen, cryo-specimen preparation, negative staining and immunogold labelling techniques, DNA and RNA tracking using hybrization in TEM or Atomic Force Microscopy, TEM crystallography and cryo TEM 3D tomography, 3D tomography of resin embedded tissues using FIB-SEM, Correlative microscopy using fluorescence microscopy, confocal microscopy or immune labelling techniques for both TEM and FIB-SEM and Elemental and isotopic identification and their distribution in cells and tissues using TEM, SEM, Scanning Transmission Electron Microscopy (STEM), Secondary Ion Mass Spectrometry (SIMS) and Nano SIMS. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Electron Microscopy: Methods and Protocols, Third Edition provides the most up-to-date and essential information in electron microscopy techniques and methods provided in this edition will assist in advancing future molecular and biological research.


Sample Preparation Handbook for Transmission Electron Microscopy

Sample Preparation Handbook for Transmission Electron Microscopy
Author: Jeanne Ayache
Publisher: Springer Science & Business Media
Total Pages: 338
Release: 2010-06-08
Genre: Technology & Engineering
ISBN: 9781441959744

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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.


Methods of Preparation for Electron Microscopy

Methods of Preparation for Electron Microscopy
Author: David G. Robinson
Publisher: Springer Science & Business Media
Total Pages: 204
Release: 2012-12-06
Genre: Science
ISBN: 364248848X

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In 1939, when the electron optics laboratory of Siemens & Halske Inc. began to manufacture the first electron microscopes, the biological and medical profes sions had an unexpected instrument at their disposal which exceeded the reso lution of the light microscope by more than a hundredfold. The immediate and broad application of this new tool was complicated by the overwhelming prob lems inherent in specimen preparation for the investigation of cellular struc tures. The microtechniques applied in light microscopy were no longer appli cable, since even the thinnest paraffin layers could not be penetrated by electrons. Many competent biological and medical research workers expressed their anxiety that objects in high vacuum would be modified due to complete dehydration and the absorbed electron energy would eventually cause degrada tion to rudimentary carbon backbones. It also seemed questionable as to whether it would be possible to prepare thin sections of approximately 0. 5 11m from heterogeneous biological specimens. Thus one was suddenly in posses sion of a completely unique instrument which, when compared with the light microscope, allowed a 10-100-fold higher resolution, yet a suitable preparation methodology was lacking. This sceptical attitude towards the application of electron microscopy in bi ology and medicine was supported simultaneously by the general opinion of colloid chemists, who postulated that in the submicroscopic region of living structures no stable building blocks existed which could be revealed with this apparatus.


Sample Preparation Handbook for Transmission Electron Microscopy

Sample Preparation Handbook for Transmission Electron Microscopy
Author: Jeanne Ayache
Publisher: Springer Science & Business Media
Total Pages: 250
Release: 2010-07-03
Genre: Technology & Engineering
ISBN: 9780387981826

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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 spe- men 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.


Electron Microscopy

Electron Microscopy
Author: John J. Bozzola
Publisher: Jones & Bartlett Learning
Total Pages: 702
Release: 1999
Genre: Medical
ISBN: 9780763701925

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New edition of an introductory reference that covers all of the important aspects of electron microscopy from a biological perspective, including theory of scanning and transmission; specimen preparation; darkroom, digital imaging, and image analysis; laboratory safety; interpretation of images; and an atlas of ultrastructure. Generously illustrated with bandw line drawings and photographs. Annotation copyrighted by Book News, Inc., Portland, OR


Specimen Preparation for Transmission Electron Microscopy of Materials

Specimen Preparation for Transmission Electron Microscopy of Materials
Author: PJ Goodhew
Publisher: Garland Science
Total Pages: 50
Release: 2020-11-25
Genre: Science
ISBN: 1000142760

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Details the essential practical steps which must precede microscopy. Methods for preparing sheet or disc specimens and final thinning techniques are described with reference to practical problems. The book also covers methods for mounting specimens in the