Methods in Visualization – serie

Although the polymerase chain reaction has revolutionized genetic analysis by amplifying rare nucleic acid sequences, the in situ application is the only method that allows the localization of amplified signal within tissue structure. The applications of in situ polymerase chain reaction have greatly enhanced the field of investigation in many disciplines, including viral infections, gene modification, tumor diagnosis, gene therapy, and cellular distribution of rare mRNA copies.PCR/RT-PCR in situ: Light and Electron Microscopy covers methods of in situ polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR), two new approaches in visualizing very low amounts of DNA and RNA in tissues and cell cultures at the light and electron microscopy levels. Written by experts in this field, the book provides theoretical consideration, as well as practical approaches to in situ PCR. The authors provide detailed protocols for each step, including the preparation of tissue samples, the rationale for the design of primers and revelation. They also emphasize the need for appropriate controls to meet the requirements of in situ PCR and RT-PCR specificity. Organized in a user-friendly two-column format, this book will provide you with tools necessary to perform and optimize these sensitive and powerful techniques in your research protocols.

Radioligand binding in situ is the most sensitive method for detecting available receptors; the success and reliability of this approach are determined by a number of technical requirements and constraints. Based on the authors' fifteen years of experience in the field, Visualization of Receptors In Situ: Applications of Radioligand Binding explicitly details these procedures - often not encountered in the original research papers - helping the reader overcome the challenges associated with this powerful technique.

In Situ hybridization allows the visualization of specific DNA/RNA sequences in individual cells in tissue sections, single cells, or chromosome preparations, and is an especially important method for studying DNA and RNA in heterogeneous cell populations. This book delves into in situ hybridization methods through the use of light microscopy used by molecular biologists, pathologists, geneticists, and biochemists. It will also appeal to research scientists who are interested in visualizing methods for nucleic acids and proteins. The book features a two-column layout for protocols/principles, simplifies the techniques and offers many schemes and tables to help the reader choose the best procedure.

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.

Histochemistry and cytochemistry are important fields for studying the inner workings of cells and tissues of the body. While visualization techniques have been in use for many years, new methods of detection developed recently help researchers and practitioners better understand cell activity. Histochemical and Cytochemical Methods of Visualization describes the essential techniques that can be used for histochemical investigations in both light and transmission electron microscopy. The book begins by discussing techniques in light microscopy. It reviews classical methods of visualization, histochemical and histoenzymatic methods, and methods used to visualize cell proliferation and apoptosis. Next, the book examines the cytochemical methods used in electron microscopy with traditional techniques, as well as more specialized methods. The final section provides an overview of image analysis and describes how image processing methods can be used to extract vital information. A 16-page insert supplies color illustrations to enhance the text.Techniques will continue to adapt to the latest technological innovations, allowing more and more precise quantification of images. These developments are essential to the biological as well as the medical sciences. This manual is a critical resource for novice and experienced researchers, technicians, and students who need to visualize what happens in the cell, the molecules expressed, the main enzymatic activities, and the repercussions of the molecular activities upon the structure of the cells in the body.