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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
Brings a fresh point of view to the current state of correlative imaging and the future of the field This book provides contributions from international experts on correlative imaging, describing their vision of future developments in the field based on where it is today. Starting with a brief historical overview of how the field evolved, it presents the latest developments in microscopy that facilitate the correlative workflow. It also discusses the need for an ideal correlative probe, applications in proteomic and elemental analysis, interpretation methods, and how correlative imaging can incorporate force microscopy, soft x-ray tomography, and volume electron microscopy techniques. Work on placing individual molecules within cells is also featured. Correlative Imaging: Focusing on the Future offers in-depth chapters on: correlative imaging from an LM perspective; the importance of sample processing for correlative imaging; correlative light and volume EM; correlation with scanning probe microscopies; and integrated microscopy. It looks at: cryo-correlative microscopy; correlative cryo soft X-ray imaging; and array tomography. Hydrated-state correlative imaging in vacuo, correlating data from different imaging modalities, and big data in correlative imaging are also considered. Brings a fresh view to one of the hottest topics within the imaging community: the correlative imaging field Discusses current research and offers expert thoughts on the field’s future developments Presented by internationally-recognized editors and contributors with extensive experience in research and applications Of interest to scientists working in the fields of imaging, structural biology, cell biology, developmental biology, neurobiology, cancer biology, infection and immunity, biomaterials and biomedicine Part of the Wiley–Royal Microscopical Society series Correlative Imaging: Focusing on the Future will appeal to those working in the expanding field of the biosciences, correlative microscopy and related microscopic areas. It will also benefit graduate students working in microscopy, as well as anyone working in the microscopy imaging field in biomedical research.
Correlative Microscopy in Biology: Instrumentation and Methods presents the detailed methodology of biological correlative microscopy, a technology that allows the acquisition of multiple data from single tissue block, cell, or section. The chapters in the book include detailed and complete instructions on the preparatory procedures. The book has 20 chapters that deal with various forms and systems of microscopy. Some of the forms and methods used in the book include light, scanning electron, fluorescence, scanning transmission electron, and ion microscopy, as well as combined light and electron and transmission electron microscope. Other methods and their applications are all discussed in detail in the book. This book will help students apply the methods without outside help as each methodology is presented in a step-by-step approach, including applications and techniques. Aside from students, the book will also be good reference for teachers, scientists, and researchers in the fields of biology, biochemistry, and medicine.
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.
Correlative Light and Electron Microscopy III, Volume 140, a new volume in the Methods in Cell Biology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Topics discussed in this new release include Millisecond time-resolved CLEM, Super resolution LM und SEM of high-pressure frozen C. elegans, Preservation fluorescence, super res CLEM, APEX in Tissue, Corrsight mit IBIDI flowthrough chamber, Correlative Light Atomic Force Electronic Microscopy (CLAFEM), Atmospheric EM CLEM, and High-precision correlation, amongst other topics. Chapters in this ongoing series deal with different approaches for analyzing the same specimen using more than one imaging technique. The strengths and application area of each is presented, with this volume exploring the aspects of sample preparation of diverse biological systems for different CLEM approaches. Contains contributions from experts in the field Covered topics include targeted ultramicrotomy and high-precision correlation Presents recent advances and currently applied correlative approaches Gives detailed protocols allowing the application of workflows in one’s own laboratory setting Covers CLEM approaches in the context of specific applications Aims to stimulate the use of new combinations of imaging modalities
Recent advances in the imaging technique electron microscopy (EM) have improved the method, making it more reliable and rewarding, particularly in its description of three-dimensional detail. Cellular Electron Microscopy will help biologists from many disciplines understand modern EM and the value it might bring to their own work. The book's five sections deal with all major issues in EM of cells: specimen preparation, imaging in 3-D, imaging and understanding frozen-hydrated samples, labeling macromolecules, and analyzing EM data. Each chapter was written by scientists who are among the best in their field, and some chapters provide multiple points of view on the issues they discuss. Each section of the book is preceded by an introduction, which should help newcomers understand the subject. The book shows why many biologists believe that modern EM will forge the link between light microscopy of live cells and atomic resolution studies of isolated macromolecules, helping us toward the goal of an atomic resolution understanding of living systems. - Updates the numerous technological innovations that have improved the capabilities of electron microscopy - Provides timely coverage of the subject given the significant rise in the number of biologists using light microscopy to answer their questions and the natural limitations of this kind of imaging - Chapters include a balance of "how to", "so what" and "where next", providing the reader with both practical information, which is necessary to use these methods, and a sense of where the field is going
Correlative Light and Electron Microscopy IV, Volume 162, a new volume in the Methods in Cell Biology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Besides the detailed description of protocols for CLEM technologies including time-resolution, Super resolution LM and Volume EM, new chapters cover Workflow (dis)-advantages/spiderweb, Serial section LM + EM, Platinum clusters as CLEM probes, Correlative Light Electron Microscopy with a transition metal complex as a single probe, SEM-TEM-SIMS, HPF-CLEM, A new workflow for high-throughput screening of mitotic mammalian cells for electron microscopy using classic histological dyes, and more. - Contains contributions from experts in the field - Covers topics using nano-SIMS and EDX for CLEM - Presents recent advances and currently applied correlative approaches - Gives detailed protocols, allowing for the application of workflows in one's own laboratory setting - Covers CLEM approaches in the context of specific applications - Aims to stimulate the use of new combinations of imaging modalities
A guide to modern scanning electron microscopy instrumentation, methodology and techniques, highlighting novel applications to cell and molecular biology.
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
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.