Immunoelectron microscopy is a key technique that bridges the information gap between biochemistry, molecular biology, and ultrastructural studies placing macromolecular functions within a cellular context. In Immunoelectron Microscopy: Methods and Protocols, expert researchers combine the tools of the molecular biologist with those of the microscopist. From the molecular biology toolbox, this volume presents methods for antigen production by protein expression in bacterial cells, methods for epitope tagged protein expression in plant and animal cells allowing protein localization in the absence of protein specific antibodies as well as methods for the production of anti-peptide, monoclonal, and polyclonal antibodies. From the microscopy toolbox, sample preparation methods for cells, plant, and animal tissue are presented. Both cryo-methods, which have the advantage of retaining protein antigenicity at the expense of ultrastructural integrity, as well as chemical fixation methods that maintain structural integrity while sacrificing protein antigenicity have been included, with chapters examining various aspects of immunogold labeling. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and essential, Immunoelectron Microscopy: Methods and Protocols seeks to facilitate an increased understanding of structure function relationships.

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).

Mitochondrial Genomics and Proteomics Protocols offers a broad collection of methods for studying the molecular biology, function, and features of mitochondria. In the past decade, mitochondrial research has elucidated the important influence of mitochondrial processes on integral cell processes such as apoptosis and cellular aging. This practical guide presents a wide spectrum of mitochondrial methods, each written by specialists with solid experience and intended for implementation by novice and expert researchers alike. Part I introduces major experimental model systems and discusses their specific advantages and limitations for functional analysis of mitochondria. The concise overview of general properties of mitochondrial systems is supplemented by detailed protocols for cultivation of model organisms. Parts II-VI comprise a robust collection of protocols for studying different molecular aspects of mitochondrial functions including: genetics and microbiology, biochemistry, physiology, dynamics and morphology, and functional genomics. Emphasis is placed on new and emerging topics in mitochondrial study, such as the examination of apoptotic effects, fusion and fission of mitochondria, and proteome and transcriptome analysis.

For this new edition, the chapters on photography and the electron microscope have been completely rewritten and two new chapters have been added--on immuno electron microscopy using colloidal gold and on useful specialized techniques.

This comprehensive reference illustrates optimal preparation methods in biological electron microscopy compared with common methodological problems. Not only will the basic methodologies of transmission electron microscopy like fixation, microtomy, and microscopy be presented, but the authors also endeavor to illustrate more specialized techniques such as negative staining, autoradiography, cytochemistry, immunoelectron microscopy, and computer-assisted image analysis. - Authored by the key leaders in the biological electron microscopy field - Illustrates both optimal and suboptimal or artifactual results in a variety of electron microscopy disciplines - Introduces students on how to read and interpret electron micrographs

This fully updated volume assembles a comprehensive collection of methods, techniques, and strategies to investigate the molecular and cellular biology of peroxisomes in different organisms. Peroxisome research is on the rise, as novel functions and proteins of this dynamic organelle are still being discovered through studies in model systems including humans, mice, flies, plants, fungi, and yeast, and this progress is reflected in the chapters included in this collection. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Peroxisomes: Methods and Protocols, Second Edition serves as an ideal guide for researchers working on peroxisome- and organelle-based research questions.

Electron microscopy in the biological sciences can be divided into two disciplines. The first, concerned with high resolution detail of particles or periodic structures, is mostly based on sound theoretical principles of physics. The second, by far the larger discipline, is interested in the information obtainable from thin sections. The theoretical back ground to those groups of techniques for preparing and looking at thin sections is often inexact and "loose", for want of a better word. What should be chemistry is often closer to alchemy. This kind of electron microscopy is often enshrined with mystical recipes, handed down from generation to generation. Admittedly, many of the processes involved, such as those required to embed tissue in epoxy resins, involve multiple interconnected steps, which make it difficult to follow the details of anyone of these steps. If all these steps are shrouded in some mystery, however, can one really trust the final image that emerges on the EM screen? When we present the data in some semi quantitative form is there really no better way to do it than to categorize the parameters with ++, +/-, etc? What happens when one labels the sections with antibodies? Does the whole business necess arily need to be more of an "art" than a "science"? Upon reflecting on these problems in 1981, I had the impression that many of the multi-authored textbooks that existed then (and that have appeared since) tended to exacerbate or at least perpetuate this

Plants, fungi, and viruses were among the first biological objects studied with an electron microscope. One of the two first instruments built by Siemens was used by Helmut Ruska, a brother of Ernst Ruska, the pioneer in constructing electron microscopes. H. Ruska published numerous papers on different biological objects in 1939. In one of these, the pictures by G. A. Kausche, E. Pfankuch, and H. Ruska of tobacco mosaic virus opened a new age in microscopy. The main problem was then as it still is today, to obtain an appropriate preparation of the specimen for observation in the electron microscope. Beam damage and specimen thickness were the first obstacles to be met. L. Marton in Brussels not only built his own instrument, but also made considerable progress in specimen preparation by introducing the impregnation of samples with heavy metals to obtain useful contrast. His pictures of the bird nest orchid root impregnated with osmium were revolutionary when published in 1934. It is not the place here to recall the different techniques which were developed in the subsequent years to attain the modern knowledge on the fine structure of plant cells and of different plant pathogens. The tremendous progress obtained with tobacco mosaic virus is reflected in the chapter by M. Wurtz on the fine structure of viruses in this Volume. New cytochemical and immunological techniques considerably surpass the morphological information obtained from the pathogens, especially at the host-parasite interface.

A mainstay for pathology residents, Autopsy Pathology is designed with a uniquely combined manual and atlas format that presents today's most complete coverage of performing, interpreting, and reporting post-mortem examinations. This lasting and useful medical reference book offers a practical, step-by-step approach to discussing not only the basics of the specialty, but the performance of specialized autopsy procedures as well. Material is divided into two sections for ease of use: a manual covering specific autopsy procedures, biosafety, generation of autopsy reports, preparation of death certificates, and other essential subjects; and an atlas, organized by organ system, which captures the appearance of the complete spectrum of autopsy findings. Offers expanded coverage of microscopic anatomy. Includes a chapter on performing special dissection procedures that may not be covered during a typical residency. Examines important techniques, such as autopsy photography and radiology, microscopic examination, supplemental laboratory studies, and other investigative approaches. Addresses the latest legal, social, and ethical issues relating to autopsies, as well as quality improvement and assurance. Presents more than 600 full-color photographs depicting common gross and microscopic autopsy findings for every part of the body. Correlates pathologic findings with their clinical causes to enhance diagnostic accuracy. Improved images in the Atlas section provide greater visual understanding. Additional online features include dissection videos demonstrating autopsy techniques; downloadable, commonly used forms for autopsy reports; and calculators for weights and measures. Expert Consult eBook version included with purchase. This enhanced eBook experience offers access to all of the text, figures, images, videos, forms, calculators, and references from the book on a variety of devices.

Since antibodies tagged with markers have been developed, immunocytochemistry has become an important method for identifying tissue substances and the localisation of nucleic acid in tissue by in situ hybridisation in molecular biology. This laboratory book covers the embedding of tissue using less sensitive epoxy resin methods to the more sensitive procedures employing the acrylics. The possibilities are discussed and results are presented so that an understanding of the techniques can be acquired.