Light And Video Microscopy

The purpose of this book is to provide the most comprehensive, easy-to-use, and informative guide on light microscopy. Light and Video Microscopy will prepare the reader for the accurate interpretation of an image and understanding of the living cell. With the presentation of geometrical optics, it will assist the reader in understanding image formation and light movement within the microscope. It also provides an explanation of the basic modes of light microscopy and the components of modern electronic imaging systems and guides the reader in determining the physicochemical information of living and developing cells, which influence interpretation. Brings together mathematics, physics, and biology to provide a broad and deep understanding of the light microscope Clearly develops all ideas from historical and logical foundations Laboratory exercises included to assist the reader with practical applications Microscope discussions include: bright field microscope, dark field microscope, oblique illumination, phase-contrast microscope, photomicrography, fluorescence microscope, polarization microscope, interference microscope, differential interference microscope, and modulation contrast microscope

Ever since television became practical in the early 1950s, closed-circuit television (CCTV) in conjunction with the light microscope has provided large screen display, raised image contrast, and made the images formed by ultraviolet and infrared rays visible. With the introduction of large-scale integrated circuits in the last decade, TV equipment has improved by leaps and bounds, as has its application in microscopy. With modem CCTV, sometimes with the help of digital computers, we can distill the image from a scene that appears to be nothing but noise; capture fluorescence too dim to be seen; visualize structures far below the limit of resolution; crispen images hidden in fog; measure, count, and sort objects; and record in time-lapsed and high-speed sequences through the light microscope without great difficulty. In fact, video is becoming indispensable for harnessing the fullest capacity of the light microscope, a capacity that itself is much greater than could have been envisioned just a few years ago. The time seemed ripe then to review the basics of video, and of microscopy, and to examine how the two could best be combined to accomplish these tasks. The Marine Biological Laboratory short courses on Analytical and Quantitative Light Microscopy in Biology, Medicine, and the Materials Sciences, and the many inquiries I received on video microscopy, supported such an effort, and Kirk Jensen of Plenum Press persuaded me of its worth.

Video microscopy is used extensively in many life and biomedical science disciplines today, and is a useful tool for both cell biologists and students. This book presents how to track the dynamic changes that take place in the structure of living cells and in reconstituted preparations using video and digital imaging microscopy. Basic information, principles, and applications are also covered, as well as more specialized video microscopy techniques. Practical laboratory guide for methods and technologies used with video microscopy Comprehensive, easy-to-follow instructions February 1998, c. 334 pp.

Ever since television became practical in the early 1950s, closed-circuit television (CCTV) in conjunction with the light microscope has provided large screen display, raised image contrast, and made the images formed by ultraviolet and infrared rays visible. With the introduction of large-scale integrated circuits in the last decade, TV equipment has improved by leaps and bounds, as has its application in microscopy. With modem CCTV, sometimes with the help of digital computers, we can distill the image from a scene that appears to be nothing but noise; capture fluorescence too dim to be seen; visualize structures far below the limit of resolution; crispen images hidden in fog; measure, count, and sort objects; and record in time-lapsed and high-speed sequences through the light microscope without great difficulty. In fact, video is becoming indispensable for harnessing the fullest capacity of the light microscope, a capacity that itself is much greater than could have been envisioned just a few years ago. The time seemed ripe then to review the basics of video, and of microscopy, and to examine how the two could best be combined to accomplish these tasks. The Marine Biological Laboratory short courses on Analytical and Quantitative Light Microscopy in Biology, Medicine, and the Materials Sciences, and the many inquiries I received on video microscopy, supported such an effort, and Kirk Jensen of Plenum Press persuaded me of its worth.

The current edition of Video Microscopy has been totally revised to reflect the advances in the tools for electronic imaging, processing, recording, and analysis, as well as applications that are being made in video microscopy and related modes of electronic imaging. The less spiral organization of the revised edition reflects an audience expected to be more experi enced in video and computer image processing than in the earlier days when coupling of video equipment and computers to the light microscope was still considered to be a novelty. Nevertheless, we have emphasi2;ed the how-tos, as well as the fundamental principles, involved in imaging and processing ih the light microscope, the human visual system, video and related electronic imaging, and the digital image processor in the hope that the reader will develop enough understanding, not only to apply rationally what is available, but also to contribute actively to the development of this evolving field. In the text, the terms appearing in the Glossary are printed in bold type, generally, at first appearance. Italics are used for emphasis.

The compound optical microscope, in its various modern forms, is probably the most familiar of all laboratory instruments and the electron microscope, once an exotic rarity, has now become a standard tool in biological and materials research. Both instruments are often used effectively with little knowledge of the relevant theory, or even of how a particular type of microscope functions. Eventually however, proper use, interpretation of images and choices of specific applications demand an understanding of fundamental principles. This book describes the principles of operation of each type of microscope currently available and of use to biomedical and materials scientists. It explains the mechanisms of image formation, contrast and its enhancement, accounts for ultimate limits on the size of observable details (resolving power and resolution) and finally provides an account of Fourier optical theory. Principles behind the photographic methods used in microscopy are also described and there is some discussion of image processing methods. The book will appeal to graduate students and researchers in the biomedical sciences, and it will be helpful to students taking a course involving the principles of microscopy.

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.

Since the first edition of Light Microscopy in Biology: A Practical approach was published, techniques in modern light microscopy have improved considerably. This fully updated edition includes revised topics from the first edition as well as coverage of techniques and technologies that have been developed since it was published. As before, the book starts with an explanation of the basic techniques, and goes on to describe current methods in: chromosome microscopy, immunohistochemistry, fluorescence microscopy, image building and video microscopy. Totally new topics covered include: confocal microscopy, calcium and pH imaging, microinjection techniques and nanovid microscopy. There are also whole chapters now devoted to reflection contrast microscopy and histomorphometry. This new edition will be of great interest to postgraduate and postdoctoral researchers in biomedicine and cell biology - both those experienced with light microscopic techniques and newcomers to the field.