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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.
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.
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 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).
This book was developed with the goal of providing an easily understood text for those users of the scanning electron microscope (SEM) who have little or no background in the area. The SEM is routinely used to study the surface structure and chemistry of a wide range of biological and synthetic materials at the micrometer to nanometer scale. Ease-of-use, typically facile sample preparation, and straightforward image interpretation, combined with high resolution, high depth of field, and the ability to undertake microchemical and crystallographic analysis, has made scanning electron microscopy one of the most powerful and versatile techniques for characterization today. Indeed, the SEM is a vital tool for the characterization of nanostructured materials and the development of nanotechnology. However, its wide use by professionals with diverse technical backgrounds—including life science, materials science, engineering, forensics, mineralogy, etc., and in various sectors of government, industry, and academia—emphasizes the need for an introductory text providing the basics of effective SEM imaging.A Beginners’ Guide to Scanning Electron Microscopy explains instrumentation, operation, image interpretation and sample preparation in a wide ranging yet succinct and practical text, treating the essential theory of specimen-beam interaction and image formation in a manner that can be effortlessly comprehended by the novice SEM user. This book provides a concise and accessible introduction to the essentials of SEM includes a large number of illustrations specifically chosen to aid readers' understanding of key concepts highlights recent advances in instrumentation, imaging and sample preparation techniques offers examples drawn from a variety of applications that appeal to professionals from diverse backgrounds.
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
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.
In this practical text, the author covers the fundamentals of biological electron microscopy - including fixation, instrumentation, and darkroom work - to provide an excellent introduction to the subject for the advanced undergraduate or graduate student.
This book provides in depths information on different microscopy approaches and supplies the reader with methods how to untangle highly complex processes involved in physiological and pathophysiological cardiac signaling. Microscopy approaches have established themselves as the quasi gold standard that enables us to appreciate the underlying mechanisms of physiological and pathophysiological cardiac signaling. This book presents the most important microscopy techniques from the level of individual molecule e.g. Förster-Resonance Energy Transfer (FRET), up to cellular and tissue imaging, e.g. electron microscopy (TEM) or light sheet microscopy. The book is intended for graduate students and postdocs in cardiovascular research, imaging and cell biology, pre-clinical and clinical researchers in cardiovascular sciences as well as decision makers of the pharmaceutical industry.
Diagnostic Electron Microscopy Diagnostic Electron Microscopy: A Practical Guide to Interpretation and Technique summarises the current interpretational applications of TEM in diagnostic pathology. This concise and accessible volume provides a working guide to the main, or most useful, applications of the technique including practical topics of concern to laboratory scientists, brief guides to traditional tissue and microbiological preparation techniques, microwave processing, digital imaging and measurement uncertainty. The text features both a screening and interpretational guide for TEM diagnostic applications and current TEM diagnostic tissue preparation methods pertinent to all clinical electron microscope units worldwide. Containing high-quality representative images, this up-to-date text includes detailed information on the most important diagnostic applications of transmission electron microscopy as well as instructions for specific tissues and current basic preparative techniques. The book is relevant to trainee pathologists and practising pathologists who are expected to understand and evaluate/screen tissues by TEM. In addition, technical and scientific staff involved in tissue preparation and diagnostic tissue evaluation/screening by TEM will find this text useful.