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Hands-on experimentalists describe the cutting-edge microscopical methods needed for the effective study of plant cell biology today. These powerful techniques, all described in great detail to ensure successful experimental results, range from light microscope cytochemistry, autoradiography, and immunocytochemistry, to recent developments in fluorescence, confocal, and dark-field microscopies. Important advances in both conventional and scanning electron microscopies are also fully developed, together with such state-of-the-art ancillary techniques as high-resolution autoradiography, immunoelectron microscopy, X-ray microanalysis, and electron systems imaging. Easy-to-use and up-to-date, Methods in Plant Electron Microscopy and Cytochemistry offers today's plant scientists a first class collection of readily reproducible light and electron microscopical methods that will prove the new standard for all working in the field.
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.
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
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.
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.
The combination of electron microscopy with transmitted light microscopy (termed correlative light and electron microscopy; CLEM) has been employed for decades to generate molecular identification that can be visualized by a dark, electron-dense precipitate. This new volume of Methods in Cell Biology covers many areas of CLEM, including a brief history and overview on CLEM methods, imaging of intermediate stages of meiotic spindle assembly in C. elegans embryos using CLEM, and capturing endocytic segregation events with HPF-CLEM. Covers many areas of CLEM by the best international scientists in the field Includes a brief history and overview on CLEM methods
With improved microscope and preparation techniques, studies of histo logical structures of plant organisms experienced a revival of interest at the end of the 19th century. From that time, histological data have sub stantially studies of the pioneers in botanical science. From the beginning of the 20th century, the microscope allowed research in cell structure, the general functional unit of living beings. Advances in cytology gradually influenced histology, at first, however, rather timidly. Only the new and spectacular progress in ultrastructural cytology and cytochemistry led to a great increase in modern work on the structures of vascular plants and the related ontogenical and physiological data, thanks to the use of the electron microscope and the contribution of molecular biology. Not only did new techniques lead to new approaches, but achieve ments in general biology shifted the orientation of research, linking in vestigation to the physiological aspects of cell and tissue differentiation. Among these, the demonstration of the general principles of develop ment, and the characterization of molecules common to plants and animals, which control and govern the main basic functions of cells and tissues, have widened the scope of modern research on plant struc tures. Present trends in biological research show that it is necessary to know the structures thoroughly, from the ultrastructural cytological scale to the scale of tissue and organ arrangement, even for physiological research on either cells, tissues, or whole organs. The study of growth factors, differ entiation, or organogenesis can be mentioned as an example.
This new edition of Plant Cell Biology balances established techniques, including classical histochemistry and electron microscopy with new developments in the field.