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first glance, it seems as simple as it is attractive: the dynamics of life are frozen in, nothing is added and nothing withdrawn except thermal energy.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.
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 go‐to resource for microscopists on biological applications of field emission gun scanning electron microscopy (FEGSEM) The evolution of scanning electron microscopy technologies and capability over the past few years has revolutionized the biological imaging capabilities of the microscope—giving it the capability to examine surface structures of cellular membranes to reveal the organization of individual proteins across a membrane bilayer and the arrangement of cell cytoskeleton at a nm scale. Most notable are their improvements for field emission scanning electron microscopy (FEGSEM), which when combined with cryo-preparation techniques, has provided insight into a wide range of biological questions including the functionality of bacteria and viruses. This full-colour, must-have book for microscopists traces the development of the biological field emission scanning electron microscopy (FEGSEM) and highlights its current value in biological research as well as its future worth. Biological Field Emission Scanning Electron Microscopy highlights the present capability of the technique and informs the wider biological science community of its application in basic biological research. Starting with the theory and history of FEGSEM, the book offers chapters covering: operation (strengths and weakness, sample selection, handling, limitations, and preparation); Commercial developments and principals from the major FEGSEM manufacturers (Thermo Scientific, JEOL, HITACHI, ZEISS, Tescan); technical developments essential to bioFEGSEM; cryobio FEGSEM; cryo-FIB; FEGSEM digital-tomography; array tomography; public health research; mammalian cells and tissues; digital challenges (image collection, storage, and automated data analysis); and more. Examines the creation of the biological field emission gun scanning electron microscopy (FEGSEM) and discusses its benefits to the biological research community and future value Provides insight into the design and development philosophy behind current instrument manufacturers Covers sample handling, applications, and key supporting techniques Focuses on the biological applications of field emission gun scanning electron microscopy (FEGSEM), covering both plant and animal research Presented in full colour An important part of the Wiley-Royal Microscopical Series, Biological Field Emission Scanning Electron Microscopy is an ideal general resource for experienced academic and industrial users of electron microscopy—specifically, those with a need to understand the application, limitations, and strengths of FEGSEM.
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.
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
Biomedical Applications of Microprobe Analysis is a combination reference/laboratory manual for the use of microprobe analysis in both clinical diagnostic and research settings. Also called microchemical microscopy, microprobe analysis uses high-energy bombardment of cells and tissue, in combination with high resolution EM or confocal microscopy to provide a profile of the ion, metal, and mineral concentrations present in a sample. This allows insight into the physiology and pathophysiology of a wide variety of cells and tissues.This book describes methods for obtaining detailed information about the identity and composition of particles too small to be seen with the naked eye and describes how this information can be useful in diagnostic and biomedical research. - Up-to-date review of electron microprobe analysis - Detailed descriptions of sample preparation techniques - Recent technologies including confocal microscopy, infrared microspectroscopy, and laser raman spectroscopy - Over 100 illustrations with numerous specific applications - Contributions by world-renowned experts in the field - Brief summary of highlights precedes each chapter
A guide to modern scanning electron microscopy instrumentation, methodology and techniques, highlighting novel applications to cell and molecular biology.
This definitive work provides a comprehensive treatment of the mathematical background and working methods of three-dimensional reconstruction from tilt series. Special emphasis is placed on the problems presented by limitations of data collection in the transmission electron microscope. The book, extensively revised and updated, takes the reader from biological specimen preparation to three-dimensional images of the cell and its components.
Presenting a unique perspective on state-of-the-art physical gels, this interdisciplinary guide provides a complete, critical analysis of the field and highlights recent developments. It shows the interconnections between the key aspects of gels, from molecules and structure through to rheological and functional properties, with each chapter focusing on a different class of gel. There is also a final chapter covering innovative systems and applications, providing the information needed to understand current and future practical applications of gels in the pharmaceutical, agricultural, cosmetic, chemical and food industries. Many research teams are involved in the field of gels, including theoreticians, experimentalists and chemical engineers, but this interdisciplinary book collates and rationalises the many different points of view to provide a clear understanding of these complex systems for researchers and graduate students.
Containing 57 thoroughly class-tested and easily customizable exercises,Laboratory Experiements in Microbiology: Tenth Edition provides engaging labs with instruction on performing basic microbiology techniques and applications for undergraduate students in diverse areas, including the biological sciences, the allied health sciences, agriculture, environmental science, nutrition, pharmacy, and various pre-professional programs. The Tenth Edition features an updated art program and a full-color design, integrating valuable micrographs throughout each exercise. Additionally, many of the illustrations have been re-rendered in a modern, realistic, three-dimensional style to better visually engage students. Laboratory Reports for each exercise have been enhanced with new Clinical Applications questions, as well as question relating to Hypotheses or Expected Results. Experiments have been refined throughout the manual and the Tenth Edition includes an extensively revised exercise on transformation in bacteria using pGLO to introduce students to this important technique.
