Hybridization Techniques For Electron Microscopy

Hybridization Techniques for Electron Microscopy examines the use of in situ hybridization techniques, including an overview of current perspectives and future developments. The book features in situ methods for fluorescence probes and confocal scanning microscopes. Three in situ hybridization methods for electron microscopes are analyzed: the non-embedded tissue method using ultrathin frozen sections, pre-embedded method, and post-embedded method using material embedded in hydrophilic resin. Positive and negative features are discussed, and clear instructions regarding implementation of techniques are provided. Particular aspects of the techniques are examined in detail, such as preparation of tissue, pretreatment, hybridization procedures, revelation (autoradiography and immunocytology) and checking procedures, in addition to the illustration, interpretation, and discussion of methods and results. The main applications described include virus detection, chromosomal gene mapping, detection of ribosomic nucleic acid, and detection of messenger RNA in animals and plants. Hybridization Techniques for Electron Microscopy is an excellent reference for cytologists, cell biologists, histochemists, cytochemists, molecular endocrinologists, and neuroendocrinologists.

In situ hybridization is a technique that allows for the visualization of specific DNA and RNA sequences in individual cells, and is an especially important method for studying nucleic acids in heterogeneous cell populations. in situ Hybridization in Electron Microscopy reviews the three main methods developed for the ultrastructural visualization

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).

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

Specimen damage in the electron microscopy; Freeze-drying for electron microscopy; Image reconstruction of electron micrographs by using equidensite integration analysis; G-banding of chromosomes; Autoradiographic localization of DNA in nonmetabolic conditions; Optical analysis and reconstruction of images; Mirror electron microscopy; Electron microscopy of banded mammalian chromosomes; Equidensitometry: some neurobiological applications.

Electron Microscopy of Plant Cells serves as manual or reference of major modern techniques used to prepare plant material for transmission and scanning electron microscopy. There have been other books that generally discuss electron microscope methodology. This book focuses on problem areas encountered through the presence of tough cell walls and large central vacuole. It details preparative techniques for botanical specimens. Each of the nine chapters of this book covers the basic principles, useful applications, and reliable procedures used on the method of electron microscopy. Other topics discussed in each chapter include the general preparation and straining of thin sections, quantitative morphological analysis, and enzyme cytochemistry. This book also explains the immunogold labelling, rapid-freezing methods, and ambient- and low-temperature scanning electron microscopy among others. This book will be invaluable to general scientists, biologists, botanists, and students specializing in plant anatomy.

The use of the term "advanced" in the title of this book is somewhat ar bitrary and very much relative with respect to time. Many techniques which were considered at the "cutting edge" of ultrastructural methodology just a few years ago are now rou tin ely used in numerous laboratories. One could cite freeze-fracture, cryothin sectioning, or indeed most of the field of scan ning electron microscopy as concrete examples. Thus the use of the term "ad vanced techniques" must be interpreted with regard to the present state of the art, and is useful only in informing the potential reader that this volume is not a primer to be used as an initial introduction into basic biological elec tron microscopy. Many excellent volumes have filled that niche in the past few years, and it is not intended that this modest book be a complete com pendium of the field. Furthermore, any limited selection of papers on advanc ed techniques necessarily reflects the preferences and arbitrary whims of the editor, thereby excluding many equally important procedures which the knowledgeable reader will readily identify. The first volume of this series appeared approximately five years ago and illustrated techniques which were thought to represent advanced and yet ba sically morphological methods for gaining increased ultrastructural informa tion from biological specimens. The present volume, on the other hand, stresses techniques which provide specific physicochemical data on the speci mens in addition to the structural information.

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