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The imaging of small cellular components requires powerful instruments, and an entire family of equipment and techniques based on the confocal principle has been developed over the past 30 years. Such methods are commonly used by neuroscience researchers, but the majority of these users do not have a microscopy or a cell biology backgrounds and do can encounter difficulties in obtaining and interpreting results. This volume brings experts in high-resolution optical microscopy applications in neuroscience and cell biology together to document the state of the art. Outlining what is currently possible, the volume also discusses promising developments for the future and aids readers in selecting the most scientifically meaningful approach to solve their questions. Each chapter discusses instrumentation and technology in relationship to application in research. All of the common and cutting edge trends are covered - fluorescence / laser electron / nonlinear microscopy, infrared fluorescence, multiphoton imaging, tomography, FRAP, live imaging, STED, PALM/STORM, etc. - Single and multiphoton confocal microscopy, and 4-pi confocal microscopy - Obtaining nanoresolution via photoactivation localization microscopy (PALM) - Several procedures that correlate observations in optical fluorescence microscopy and electron microscopy - Study of morphology and function via high-resolution fluorescence procedures - Additional high-resolution microscopic techniques
In the biomedical sciences, the confocal laser scanning microscope (CLSM) has become the instrument of choice for producing high-resolution images and 3D reconstruction, breaking the barriers of conventional optical microscopy. Wouterlood (anatomy, VU U. Medical Center, Amsterdam, the Netherlands) introduces the confocal principle which eliminates out-of-focus haze, its components, and relevant equations. International scientists explain the principles and related methods of stimulated emission depletion (SRED), single molecule localization, and coherent anti-Stokes Raman (CARS) microscopy; labeling approaches; preparation of samples for imaging; and applications of, and developments in, this new wave of imaging, e.g., visualization of neuronal networks, DNA, and myelin. The text includes color and b&w images, and referral to an online CLSM simulator. Academic Press is an imprint of Elsevier. Annotation ©2013 Book News, Inc., Portland, OR (booknews.com).
As imaging studies have continued to expand in scope and sophistication, this new edition of the highly successful and well–received Imaging Neurons: A Laboratory Manualhas expanded to include development, with over twenty new chapters on such topics as MRI microscopy, imaging early developmental events, and labeling single neurons. Chapters on FRET, FCS/ICS, FRAP, hyperresolution microscopy, single molecule imaging, imaging with quantum dots, and imaging gene expression are included. With over forty full chapters, the manual also includes over forty sections of protocols for imaging techniques.
In the past decade, advances in microscopy have been coupled with new methods of culturing and labeling cells to generate the new science of imaging. Imaging technologies allow investigators to look directly inside living cells and probe their form and function in unprecedented detail. This approach is revolutionizing many aspects of biomedical research, particularly neuroscience, in which visual techniques have traditionally been so important. This manual is the first comprehensive description of the range of imaging technologies being applied to living cells. With its origins in a laboratory course taught at Cold Spring Harbor Laboratory by the editors and contributors, it is packed with the kind of technical detail and practical advice that are essential for success, yet seldom found in the research literature. It covers both established methods and cutting-edge techniques such as multiphoton excitation microscopy and imaging of genetically engineered probes. Although it is neurons to which these technologies are most commonly applied, the methods described are readily adaptable to many other cell types. This book will therefore be an invaluable aid to investigators in cell and developmental biology and immunology as well as neuroscience who wish to take advantage of the extraordinary insights into cellular function offered by imaging technologies.
The Handbook of Neurophotonics provides a dedicated overview of neurophotonics, covering the use of advanced optical technologies to record, stimulate, and control the activity of the brain, yielding new insight and advantages over conventional tools due to the adaptability and non-invasive nature of light. Including 32 colour figures, this book addresses functional studies of neurovascular signaling, metabolism, electrical excitation, and hemodynamics, as well as clinical applications for imaging and manipulating brain structure and function. The unifying theme throughout is not only to highlight the technology, but to show how these novel methods are becoming critical to breakthroughs that will lead to advances in our ability to manage and treat human diseases of the brain. Key Features: Provides the first dedicated book on state-of-the-art optical techniques for sensing and imaging across at the cellular, molecular, network, and whole brain levels. Highlights how the methods are used for measurement, control, and tracking of molecular events in live neuronal cells, both in basic research and clinical practice. Covers the entire spectrum of approaches, from optogenetics to functional methods, photostimulation, optical dissection, multiscale imaging, microscopy, and structural imaging. Includes chapters that show use of voltage-sensitive dye imaging, hemodynamic imaging, multiphoton imaging, temporal multiplexing, multiplane microscopy, optoacoustic imaging, near-infrared spectroscopy, and miniature neuroimaging devices to track cortical brain activity.
If a picture is worth a thousand words, then dynamic images of brain activity certainly warrant many, many more. This book will help users learn to decipher the dynamic imaging data that will be critical to our future understanding of complex brain functions. In recent years, there have been unprecedented methodological advancements in the imaging of brain activity. These techniques allow the measurement of everything from neural activity (e.g., membrane potential, ion ?ux, neurotransmitter ?ux) to energy metabolism (e.g., glucose consumption, oxygen consumption, creatine kinase ?ux) and functional hyperemia (e.g., blood ?ow, volume, oxygenation). This book deals with a variety of magnetic resonance, electrophysiology, and optical methods that are often used to measure some of these dynamic processes. All chapters were written by leading experts, spanning three continents, specializing in state-of-the-art methods. Brie?y, the book has ?ve sections. In the introductory section, there are two chapters; the ?rst one contains a brief pre- ble to dynamic brain imaging and the other presents a novel, analytical approach to processing of dynamically acquired data. The second section has four chapters and delves into a wide range of optical imaging methods. I am privileged to include a chapter from Lawrence B. Cohen, considered by many to be the authority on optical imaging and spectroscopy, both in vitro and in vivo [Cohen LB (1973) Physiol Rev.
Cell Imaging is rapidly evolving as new technologies and new imaging advances continue to be introduced. In the second edition of Cell Imaging Techniques: Methods and Protocols expands upon the previous editions with current techniques that includes confocal microscopy, transmission electron microscopy, atomic force microscopy, and laser microdissection. With new chapters covering colocalization analysis of fluorescent probes, correlative light and electron microscopy, environmental scanning electron microscopy, light sheet microscopy, intravital microscopy, high throughput microscopy, and stereological techniques. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls Authoritative and cutting-edge, Cell Imaging Techniques: Methods and Protocols, Second Edition is an easily accessible volume of protocols to be used with a variety of imaging-based equipment likely available in a core imaging facility.
Modern neuroscience research is inherently multidisciplinary, with a wide variety of cutting edge new techniques to explore multiple levels of investigation. This Third Edition of Guide to Research Techniques in Neuroscience provides a comprehensive overview of classical and cutting edge methods including their utility, limitations, and how data are presented in the literature. This book can be used as an introduction to neuroscience techniques for anyone new to the field or as a reference for any neuroscientist while reading papers or attending talks. - Nearly 200 updated full-color illustrations to clearly convey the theory and practice of neuroscience methods - Expands on techniques from previous editions and covers many new techniques including in vivo calcium imaging, fiber photometry, RNA-Seq, brain spheroids, CRISPR-Cas9 genome editing, and more - Clear, straightforward explanations of each technique for anyone new to the field - A broad scope of methods, from noninvasive brain imaging in human subjects, to electrophysiology in animal models, to recombinant DNA technology in test tubes, to transfection of neurons in cell culture - Detailed recommendations on where to find protocols and other resources for specific techniques - "Walk-through" boxes that guide readers through experiments step-by-step