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In Confocal Microscopy Methods and Protocols, Stephen Paddock and a highly skilled panel of experts lead the researcher using confocal techniques from the bench top, through the imaging process, to the journal page. They concisely describe all the key stages of confocal imaging-from tissue sampling methods, through the staining process, to the manipulation, presentation, and publication of the realized image. Written in a user-friendly, nontechnical style, the methods specifically cover most of the commonly used model organisms: worms, sea urchins, flies, plants, yeast, frogs, and zebrafish. Centered in the many biological applications of the confocal microscope, the book makes possible the successful imaging of both fixed and living specimens using primarily the laser scanning confocal microscope. The powerful hands-on methods collected in Confocal Microscopy Methods and Protocols will help even the novice to produce first-class cover-quality confocal images.
This third edition of a classic text in biological microscopy includes detailed descriptions and in-depth comparisons of parts of the microscope itself, digital aspects of data acquisition and properties of fluorescent dyes, the techniques of 3D specimen preparation and the fundamental limitations, and practical complexities of quantitative confocal fluorescence imaging. Coverage includes practical multiphoton, photodamage and phototoxicity, 3D FRET, 3D microscopy correlated with micro-MNR, CARS, second and third harmonic signals, ion imaging in 3D, scanning RAMAN, plant specimens, practical 3D microscopy and correlated optical tomography.
Confocal laser microscopy is a microscopic observation technique patented by Minsk in 1957. Its success is due to the undeniable advantages over traditional optical microscopy including improved image clarity and contrast, greater vertical and horizontal resolution, etc., and above all the possibility to take optical sections of the sample, which allows us to obtain a tridimensional study. This authors of this book provide and discuss new research on the principles, techniques and applications of confocal microscopy.
There has been a great upsurge in interest in light microscopy in recent years due to the advent of a number of significant advances in microscopy, one of the most important of which is confocal microscopy. Confocal microscopy has now become an important research tool, with a large number of new fluorescent dyes becoming available in the past few years, for probing your pet structure or molecule within fixed or living cell or tissue sampies. Many of the people interested in using confocal microscopy to further their research do not have a background in microscopy or even cell biology and so not only do they find considerable difficulty in obtaining satisfactory results with a confocal microscope, but they may be mislead by how data is being presented. This book is intended to teach you the basic concepts ofmicroscopy, fluorescence, digital imaging and the principles of confocal microscopy so that you may take full advantage ofthe excellent confocal microscopes now available. This book is also an excellent reference source for information related to confocal microscopy for both beginners and the more advanced users. For example, do you need to know the optimal pinhole size for a 63x 1. 4 NA lens? Do you need to know the fluorescence emission spectrum of Alexa 568? Access to the wealth of practical information in this book is made easier by using both the detailed index and the extensive glossary.
Basic Confocal Microscopy, Second Edition builds on the successful first edition by keeping the same format and reflecting relevant changes and recent developments in this still-burgeoning field. This format is based on the Confocal Microscopy Workshop that has been taught by several of the authors for nearly 20 years and remains a popular workshop for gaining basic skills in confocal microscopy. While much of the information concerning fluorescence and confocal microscopy that made the first edition a success has not changed in the six years since the book was first published, confocal imaging is an evolving field and recent advances in detector technology, operating software, tissue preparation and clearing, image analysis, and more have been updated to reflect this. Several of these advances are now considered routine in many laboratories, and others such as super resolution techniques built on confocal technology are becoming widely available.
As part of the Reliable Lab Solutions series, Techniques in Confocal Microscopy brings together chapters from volumes 302, 307 and 356 of Methods in Enzymology. It documents many diverse uses for confocal microscopy in disciplines that broadly span biology. Documents many diverse uses for confocal microscopy in disciplines that broadly span biology The methods presented include shortcuts and conveniences not included in the initial publications Techniques are described in a context that allows comparisons to other related methodologies Methodologies are laid out in a manner that stresses their general applicability and reports their potential limitations
With contributions by numerous experts
This comprehensive reference work provides an overview of, and practical guide to, the various computer-aided microscopical techniques used in materials science today. After introducing the reader to the basic concepts of optics, the interactions between light and matter, and image processing, the book goes on to discuss in depth both 2D reflection microscopy and confocal laser scanning microscopy. The application of these techniques to the characterisation of materials is abundantly illustrated by hundreds of photographs and illustrations, and through specific case studies. There is also discussion of other modern optical imaging techniques and of non-optical ones such as x-ray micrography. This reference text is essential both for beginners looking for an introduction to the subject as well as advanced materials researchers in the fields where optical microscopy is used. Major reference work on the application of microscopy techniques to materials science research Includes over 420 photographs and illustrations Provides detailed coverage of the major light microscopical techniques including optical reflection microscopy and confocal laser scanning microscopy as well as novel techniques such raman microscopy, tomography and microtomography
A diverse collection of state-of-the-art methods for the microscopic imaging of cells and molecules. The authors cover a wide spectrum of complimentary techniques, including such methods as fluorescence microscopy, electron microscopy, atomic force microscopy, and laser scanning cytometry. Additional readily reproducible protocols on confocal scanning laser microscopy, quantitative computer-assisted image analysis, laser-capture microdissection, microarray image scanning, near-field scanning optical microscopy, and reflection contrast microscopy round out this eclectic collection of cutting-edge imaging techniques now available. The authors also discuss preparative methods for particles and cells by transmission electron microscopy.
This text guides you through the principles and practical techniques of confocal and multiphoton microscopy. It also describes the historical connections and parallel inventions that resulted in modern techniques of live cell imaging and their use in biology and medicine. You will find comparisons of different types of confocal and multiphoton microscopes, solutions to the problems one would encounter when using various microscopic techniques, tips on selecting equipment, and an extensive annotated bibliography of additional resources.