Biological Electron Microscopy

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.

Major improvements in instrumentation and specimen preparation have brought SEM to the fore as a biological imaging technique. Although this imaging technique has undergone tremendous developments, it is still poorly represented in the literature, limited to journal articles and chapters in books. This comprehensive volume is dedicated to the theory and practical applications of FESEM in biological samples. It provides a comprehensive explanation of instrumentation, applications, and protocols, and is intended to teach the reader how to operate such microscopes to obtain the best quality images.

The past decade has seen a remarkable increase in the use of electron microscopy as a researm tool in biology and medicine. Thus, most institu tions of higher learning now boast several electron optical laboratories having various levels of sophistication. Training in the routine use of elec tron optical equipment and interpretation of results is no longer restricted to a few prestigious centers. On the other hand, temniques utilized by researm workers in the ultrastructural domain have become extremely diverse and complex. Although a large number of quite excellent volumes of electron microscopic temnique are now dedicated to the basic elements available whim allow the novice to acquire a reasonable introduction to the field, relatively few books have been devoted to a discussion of more ad vanced temnical aspects of the art. It was with this view that the present volume was conceived as a handy reference for workers already having some background in the field, as an information source for those wishing to shift efforts into more promising temniques, or for use as an advanced course or seminar guide. Subject matter has been mosen particularly on the basis of pertinence to present researm activities in biological electron microscopy and emphasis has been given those areas whim seem destined to greatly expand in useful ness in the near future.

In the continuing quest to explore structure and to relate struc tural organization to functional significance, the scientist has developed a vast array of microscopes. The scanning electron microscope (SEM) represents a recent and important advance in the development of useful tools for investigating the structural organization of matter. Recent progress in both technology and methodology has resulted in numerous biological publications in which the SEM has been utilized exclusively or in connection with other types of microscopes to reveal surface as well as intracellular details in plant and animal tissues and organs. Because of the resolution and depth of focus presented in the SEM photograph when compared, for example, with that in the light microscope photographs, images recorded with the SEM have widely circulated in newspapers, periodicals and scientific journals in recent times. Considering the utility and present status of scanning electron microscopy, it seemed to us to be a particularly appropriate time to assemble a text-atlas dealing with biological applications of scanning electron microscopy so that such information might be presented to the student and to others not yet familiar with its capabilities in teaching and research. The major goal of this book, therefore, has been to assemble material that would be useful to those students beginning their study of botany or zoo logy, as well as to beginning medical students and students in advanced biology courses.

The go‐to resource for microscopists on biological applications of field emission gun scanning electron microscopy (FEGSEM) The evolution of scanning electron microscopy technologies and capability over the past few years has revolutionized the biological imaging capabilities of the microscope—giving it the capability to examine surface structures of cellular membranes to reveal the organization of individual proteins across a membrane bilayer and the arrangement of cell cytoskeleton at a nm scale. Most notable are their improvements for field emission scanning electron microscopy (FEGSEM), which when combined with cryo-preparation techniques, has provided insight into a wide range of biological questions including the functionality of bacteria and viruses. This full-colour, must-have book for microscopists traces the development of the biological field emission scanning electron microscopy (FEGSEM) and highlights its current value in biological research as well as its future worth. Biological Field Emission Scanning Electron Microscopy highlights the present capability of the technique and informs the wider biological science community of its application in basic biological research. Starting with the theory and history of FEGSEM, the book offers chapters covering: operation (strengths and weakness, sample selection, handling, limitations, and preparation); Commercial developments and principals from the major FEGSEM manufacturers (Thermo Scientific, JEOL, HITACHI, ZEISS, Tescan); technical developments essential to bioFEGSEM; cryobio FEGSEM; cryo-FIB; FEGSEM digital-tomography; array tomography; public health research; mammalian cells and tissues; digital challenges (image collection, storage, and automated data analysis); and more. Examines the creation of the biological field emission gun scanning electron microscopy (FEGSEM) and discusses its benefits to the biological research community and future value Provides insight into the design and development philosophy behind current instrument manufacturers Covers sample handling, applications, and key supporting techniques Focuses on the biological applications of field emission gun scanning electron microscopy (FEGSEM), covering both plant and animal research Presented in full colour An important part of the Wiley-Royal Microscopical Series, Biological Field Emission Scanning Electron Microscopy is an ideal general resource for experienced academic and industrial users of electron microscopy—specifically, those with a need to understand the application, limitations, and strengths of FEGSEM.

This book presents the newest technology in electron microscopy. It comprises two major areas of electron microscopy - transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The volume provides clear, concise instructions on processing biological specimens and includes discussion on the underlying principles of the majority of the processes presented. A notes section enables efficient adaptation and troubleshooting of protocols.

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.

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.