This volume discusses different approaches to workflows for large volume electron microscopy - from preparation of samples to their imaging in a variety of microscopes - in some cases also applying correlative techniques. The chapters in this book cover topics such as correlative super resolution and electron microscopy to detect molecules in their native cellular context; low-threshold access to serial section arrays; improving serial blockface SEM by focal charge compensation; FIBSEM analysis of interfaces between hard technical devices and soft neuronal tissue; and image processing for volume electron microscopy. In Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory. Cutting-edge and authoritative, Volume Microscopy: Multiscale Imaging with Photons, Electrons, and Ions is a valuable resource for novice and expert scientists interested in learning more about this evolving field.

This new volume, number 123, of Methods in Cell Biology looks at methods for quantitative imaging in cell biology. It covers both theoretical and practical aspects of using optical fluorescence microscopy and image analysis techniques for quantitative applications. The introductory chapters cover fundamental concepts and techniques important for obtaining accurate and precise quantitative data from imaging systems. These chapters address how choice of microscope, fluorophores, and digital detector impact the quality of quantitative data, and include step-by-step protocols for capturing and analyzing quantitative images. Common quantitative applications, including co-localization, ratiometric imaging, and counting molecules, are covered in detail. Practical chapters cover topics critical to getting the most out of your imaging system, from microscope maintenance to creating standardized samples for measuring resolution. Later chapters cover recent advances in quantitative imaging techniques, including super-resolution and light sheet microscopy. With cutting-edge material, this comprehensive collection is intended to guide researchers for years to come. Covers sections on model systems and functional studies, imaging-based approaches and emerging studies Chapters are written by experts in the field Cutting-edge material

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.

Scanning Electron Microscopy provides a description of the physics of electron-probe formation and of electron-specimen interactions. The different imaging and analytical modes using secondary and backscattered electrons, electron-beam-induced currents, X-ray and Auger electrons, electron channelling effects, and cathodoluminescence are discussed to evaluate specific contrasts and to obtain quantitative information.

Volume Electron Microscopy (vEM), Volume 177 is a collective term for a set of three-dimensional high-resolution ultrastructural imaging techniques that have delivered new insights into biological systems in recent years, garnering substantial interest in the life and clinical sciences. In this book, users will find a variety of vEM workflows and technologies, highlighting application areas with biologically relevant examples. Topics covered include Automated large volume sample preparation for vEM, Resin comparison for Serial Block Face Scanning Volume Electron Microscopy, Immunolabelling for SBF-SEM, Electron Microscopy in Plants, Serial section electron tomography, Automated Tape Collecting Ultramicrotomy (ATUM) for targeting neuropathology, Array tomography, and much more. Other sections focus on Mitochondria morphometry in 3d datasets of mouse brain obtained with serial block-face Scanning Electron Microscopy, Serial Block-face Scanning Electron Microscopy of Schmidtea mediterranea, Correlative multiscale microCT-SBF-SEM imaging of resin-embedded tissue, Methods of enhanced FIB-SEM sample preparation and image acquisition, Functional characterization of endo-lysosomal compartments by correlative live-cell and volume electron microscopy, and much more. - Includes chapters written by key leaders and developers in the field - Provides detailed protocols, allowing for the application of workflows in one's own laboratory setting - Presents real tips and tricks you won't get from standard research papers

The previous edition of this book marked the shift in technology from video to digital camera use with microscope use in biological science. This new edition presents some of the optical fundamentals needed to provide a quality image to the digital camera. Specifically, it covers the fundamental geometric optics of finite- and infinity-corrected microscopes, develops the concepts of physical optics and Abbe's theory of image formation, presents the principles of Kohler illumination, and finally reviews the fundamentals of fluorescence and fluorescence microscopy. The second group of chapters deals with digital and video fundamentals: how digital and video cameras work, how to coordinate cameras with microscopes, how to deal with digital data, the fundamentals of image processing, and low light level cameras. The third group of chapters address some specialized areas of microscopy that allow sophisticated measurements of events in living cells that are below the optical limits of resolution. - Expands coverage to include discussion of confocal microscopy not found in the previous edition - Includes "traps and pitfalls" as well as laboratory exercises to help illustrate methods

Expansion Microscopy for Cell Biology, Volume 161 in the Methods in Cell Biology series, compiles recent developments in expansion microscopy techniques (Pro-ExM, U-ExM, Ex-STED, X10, Ex-dSTORM, etc.) and their applications in cell biology, ranging from mitosis, centrioles or nuclear pore complex to plant cell, bacteria, Drosophila or neurons. Chapters in this new release include Protein-retention Expansion Microscopy: Improved Sub-cellular Imaging Resolution through Physical Specimen Expansion, Ultrastructure Expansion Microscopy (U-ExM), Expansion STED microscopy (ExSTED), Simple multi-color super-resolution by X10 microscopy, Expansion microscopy imaging of various neuronal structures, Mapping the neuronal cytoskeleton using expansion microscopy, Mechanical expansion microscopy, and much more. - Provides the authority and expertise of leading contributors from an international board of authors - Represents the latest release in the Methods in Cell Biology series - Includes the latest information on Expansion Microscopy for Cell Biology

Fluorescence Microscopy of Living Cells in Culture, Part B

This book has evolved by processes of selection and expansion from its predecessor, Practical Scanning Electron Microscopy (PSEM), published by Plenum Press in 1975. The interaction of the authors with students at the Short Course on Scanning Electron Microscopy and X-Ray Microanalysis held annually at Lehigh University has helped greatly in developing this textbook. The material has been chosen to provide a student with a general introduction to the techniques of scanning electron microscopy and x-ray microanalysis suitable for application in such fields as biology, geology, solid state physics, and materials science. Following the format of PSEM, this book gives the student a basic knowledge of (1) the user-controlled functions of the electron optics of the scanning electron microscope and electron microprobe, (2) the characteristics of electron-beam-sample inter actions, (3) image formation and interpretation, (4) x-ray spectrometry, and (5) quantitative x-ray microanalysis. Each of these topics has been updated and in most cases expanded over the material presented in PSEM in order to give the reader sufficient coverage to understand these topics and apply the information in the laboratory. Throughout the text, we have attempted to emphasize practical aspects of the techniques, describing those instru ment parameters which the microscopist can and must manipulate to obtain optimum information from the specimen. Certain areas in particular have been expanded in response to their increasing importance in the SEM field. Thus energy-dispersive x-ray spectrometry, which has undergone a tremendous surge in growth, is treated in substantial detail.

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).