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A book for physiologists, biologists and biochemists studying cell physiology. Included are major optical techniques and bases for designing experiments and choosing appropriate instrumentation, along with discussions on methods and results of optical techniques applied to research through image enhancement, probes for membrane potential, intracellular indicators for Ca and pH, and photobleaching and photoactivation techniques. Also included are schematic drawings and numerous references.
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
Volume 55 in Methods in Cell Biology is a concise laboratory book that emphasizes the methods and technologies needed to use single polarized laser light source that functions simultaneously as an optical trap and a dual-beam interferometer.* * Provides a practical laboratory guide for methods and technologies used with laser tweezers* Includes comprehensive and easy-to-follow protocols
Advances in technology have revolutionized the development of light microscopy techniques in biomedical research, thus improving visualization of the microstructure of cells and tissues under physiological conditions. Fluorescence microscopy methods are non-contact and non-invasive and provide high spatial and temporal resolution that other laboratory techniques cannot. This well-illustrated book targets graduate students and scientists who are new to the state-of-the-art fluorescence microscopy techniques used in biological and clinical imaging. It explains basic concepts and imaging procedures for wide-field, confocal, multiphoton excitation, fluorescence resonance energy transfer (FRET), lifetime imaging (FLIM), spectral imaging, fluorescence recovery after photobleaching (FRAP), optical tweezers, total internal reflection, high spatial resolution atomic force microscopy (AFM), and bioluminescence imaging for gene expression. The usage of these techniques in various biological applications, including calcium, pH, membrane potential, mitochondrial signaling, protein-protein interactions under various physiological conditions, and deep tissue imaging, is clearly presented. The authors describe the approaches to selecting epifluorescence microscopy, the detectors, and the image acquisition and processing software for different biological applications. Step-by-step directions on preparing different digital formats for light microscopy images on websites are also provided.
Optical Imaging Techniques in Cell Biology, Second Edition covers the field of biological microscopy, from the optics of the microscope to the latest advances in imaging below the traditional resolution limit. It includes the techniques-such as labeling by immunofluorescence and fluorescent proteins-which have revolutionized cell biology. Quantitat
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).
Optical probes, particularly the fluorescent varieties, enable researchers to observe cellular events in real time and with great spatial resolution. Optical Probes in Biology explores the diverse capabilities of these powerful and versatile tools and presents various approaches used to design, develop, and implement them. The book examines the use
Optical Microscopy: Emerging Methods and Applications covers recent technical advances and new approaches to monitoring and altering cell physiology, examining membrane cytoarchitecture, observing multiple cellular activities and intact organ physiology, plus confocal imaging of live cell function, lifetime imaging, and automated clinical imaging cytometry. The book provides the reader with a synopsis of the most recent technical developments in optical microscopy as applied to scientific research. Each chapter introduces new methods by describing how these overcome limitations inherent in previous techniques. Software, hardware, and other equipment concerns are covered. Additionally, the book reviews current applications in order to stimulate future developments in optical microscopy, encouraging novel uses and new technical advances - Caged compounds, fluorescence ratio imaging, and CCD video cameras - Simultaneous multiple detection and real-time fluorescence microscopy - Simultaneous DIC and quantitative LLF video imaging - Total internal reflectance, time-resolved, and automated fluorescence microscopy - Laser-scanning confocal microscopy - Imaging for calcium measurements, membranes, glycoproteins, living cells, and cancer cells
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
In spite of tremendous scientific progress over the past years, cell biologists do not yet understand the fundamental processes that determine the life cy cle of a cell. Such are: cell movement and cell spreading, cell division, cell communication, cell signaling, cell regeneration and cell death. Biochemistry has enabled us to recognize and to isolate an overwhelming number of new proteins. In vitro assays and the reinjection of proteins into cells and tissues have provided insights into molecular functions and cellular mechanisms. The renaissance of the genetic approach by applying restriction enzymes and vectors, PCR and antisense technology has enabled us to overexpress certain cellular products, to make altered constructs of cell components or to create "knock-out" mutants that entirely lack the factor of interest. Amazingly en ough, all these molecular toys have led to a stream of information but not, in a comparable degree, to a better understanding. Has the puzzle become too complex to get solved; or are the windows too small that we are looking through? As an attempt to answer both questions, the aim of the present mono graph Modern Optics, Electronics and High Precision Techniques in Cell Biol ogy is first to provide cell and molecular biologists with a whole new scope of easily applicable techniques including brand-new optical, biophysical, physicochemical and biosensoric devices. Secondly, these newly developed techniques allow us to look at cells and biological systems as a whole.