Biological Techniques In 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.

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.

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

Some Biological Techniques in Electron Microscopy discusses the problems that hinder or prevent progress in biological electron microscopy. This book explores the great potential of electron microscope for solving several pressing medical and biological research problems. Organized into five chapters, this book starts with an overview of the primary goal of biological electron microscopy, which is the visualization of atoms in biological molecules and structures. This text proceeds with a discussion of the freezing methods of electron microscopy in which some of the artifacts producing steps of environmental techniques are eliminated. Other chapters consider the objective of electron microscopy to resolve successively small objects, in the limiting instance, single atoms in amorphous structures. The final chapter deals with processing tissues for electron microscopy wherein temperature, agitation, pressure, and humidity can be controlled throughout the operation. Biologists, biophysicists, physicists, electron microscopists, and medical researchers will find this book extremely useful.

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).

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.

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.