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SPIE Milestones are collections of seminal papers from the world literature covering important discoveries and developments in optics and photonics.
In the last decade multiphoton excitation microscopy has emerged as an important technique with ever increasing numbers of significant applications in the fields of biology, chemistry, physics, and medicine. This volume contains key papers on the following topics: developments of nonlinear optical spectroscopy and nonlinear scanning microscopy (SHG, CARS); theory and techniques of multiphoton excitation microscopy; development of laser sources; single-molecule studies; and applications to biology, cell biology, embryology and developmental biology, neuroscience, dermatology, and optical biopsy. A comprehensive bibliography follows the reprinted papers.
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.
This book provides a comprehensive introduction to the field of scanning optical microscopy for scientists and engineers. The book concentrates mainly on two instruments: the Confocal Scanning Optical Microscope (CSOM), and the Optical Interference Microscope (OIM). A comprehensive discussion of the theory and design of the Near-Field Scanning Optical Microscope (NSOM) is also given. The text discusses the practical aspects of building a confocal scanning optical microscope or optical interference microscope, and the applications of these microscopes to phase imaging, biological imaging, and semiconductor inspection and metrology.A comprehensive theoretical discussion of the depth and transverse resolution is given with emphasis placed on the practical results of the theoretical calculations and how these can be used to help understand the operation of these microscopes. - Provides a comprehensive introduction to the field of scanning optical microscopy for scientists and engineers - Explains many practical applications of scanning optical and interference microscopy in such diverse fields as biology and semiconductor metrology - Discusses in theoretical terms the origin of the improved depth and transverse resolution of scanning optical and interference microscopes with emphasis on the practical results of the theoretical calculations - Considers the practical aspects of building a confocal scanning or interference microscope and explores some of the design tradeoffs made for microscopes used in various applications - Discusses the theory and design of near-field optical microscopes - Explains phase imaging in the scanning optical and interference microscopes
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.
Numerous applications of confocal microscopes include the life sciences, ophthamology, industrial inspection and semiconductor linewidth metrology. Concentrating on the science and applications of confocal microscopy, this book includes all the latest developments in three-dimensional processing techniques.
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.
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.
The confocal microscope is appropriate for imaging cells or the measurement of industrial artefacts. However, junior researchers and instrument users sometimes misuse imaging concepts and metrological characteristics, such as position resolution in industrial metrology and scale resolution in bio-imaging. And, metrological characteristics or influence factors in 3D measurement such as height assessment error caused by 3D coupling effect are so far not yet identified. In this book, the authors outline their practices by the working experiences on standardization and system design. This book assumes little previous knowledge of optics, but rich experience in engineering of industrial measurements, in particular with profile metrology or areal surface topography will be very helpful to understand the theoretical concerns and value of the technological advances. It should be useful for graduate students or researchers as extended reading material, as well as microscope users alongside their handbook.
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