Light and Video Microscopy

Light and Video Microscopy

2nd Edition - December 16, 2013

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  • Author: Randy Wayne
  • Hardcover ISBN: 9780124114845
  • eBook ISBN: 9780124115361

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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.

Key Features

  • 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


Cell biologists, neurobiologists, reproductive biologists, plant biologists, microbiologists, fungal biologists, veterinarians, forensic scientists, food scientists, horticulturalists, biochemists, physicists, engineers, and anyone using a light or video microscope

Table of Contents

  • Dedication

    Preface to the Second Edition

    Preface to the First Edition

    Chapter 1. The Relation Between the Object and the Image

    Luminous and Nonluminous Objects

    Object and Image

    Theories of Vision

    Light Travels in Straight Lines

    Images Formed in a Camera Obscura: Geometric Considerations

    Where Does Light Come From?

    How Can the Amount of Light Be Measured?

    Web Resources

    Chapter 2. The Geometric Relationship Between Object and Image

    Reflection by a Plane Mirror

    Reflection by a Curved Mirror

    Reflection from Various Sources

    Images Formed by Refraction at a Plane Surface

    Images Formed by Refraction at a Curved Surface

    Fermat’s Principle

    Optical Path Length

    Lens Aberrations

    Geometric Optics and Biology

    Geometric Optics of the Human Eye

    Web Resources

    Chapter 3. The Dependence of Image Formation on the Nature of Light

    Christiaan Huygens and the Invention of the Wave Theory of Light

    Thomas Young and the Development of the Wave Theory of Light

    James Clerk Maxwell and the Wave Theory of Light

    Ernst Abbe and the Relationship of Diffraction to Image Formation

    Resolving Power and the Limit of Resolution


    Web Resources

    Chapter 4. Bright-Field Microscopy

    Components of the Microscope

    The Optical Paths of the Light Microscope

    Using the Bright-Field Microscope

    Depth of Field

    Out-of-Focus Contrast

    Uses of Bright-Field Microscopy

    Care and Cleaning of the Light Microscope

    Web Resources

    Chapter 5. Photomicrography

    Setting up the Microscope for Photomicrography

    Scientific History of Photography

    General Nature of the Photographic Process

    The Resolution of the Film

    Exposure and Composition

    The Similarities between Film and the Retina

    Web Resources

    Chapter 6. Methods of Generating Contrast

    Dark-Field Microscopy

    Rheinberg Illumination

    Oblique Illumination

    Phase-Contrast Microscopy

    Oblique Illumination Reconsidered

    Annular Illumination

    Chapter 7. Polarization Microscopy

    What is Polarized Light?

    Use an Analyzer to Test for Polarized Light

    Production of Polarized Light

    Influencing Light

    Design of a Polarizing Microscope

    What is the Molecular Basis of Birefringence?

    Interference of Polarized Light

    The Origin of Colors in Birefringent Specimens

    Use of Compensators to Determine the Magnitude and Sign of Birefringence

    Crystalline versus Form Birefringence

    Orthoscopic versus Conoscopic Observations

    Reflected Light Polarization Microscopy

    Uses of Polarization Microscopy

    Optical Rotatory (or Rotary) Polarization and Optical Rotatory (or Rotary) Dispersion

    Web Resources

    Chapter 8. Interference Microscopy

    Generation of Interference Colors

    The Relationship of Interference Microscopy to Phase-Contrast Microscopy

    Quantitative Interference Microscopy: Determination of the Refractive Index, Mass, Concentration of Dry Matter, Concentration of Water, and Density

    Source of Errors When Using an Interference Microscope

    Making a Coherent Reference Beam

    Double-Beam versus Multiple-Beam Interference

    Interference Microscopes Based on a Mach–Zehnder Type Interferometer

    Interference Microscopes Based on Polarized Light

    The Use of Transmission Interference Microscopy in Biology

    Reflection–Interference Microscopy

    Uses of Reflection–Interference Microscopy in Biology

    Web Resources

    Chapter 9. Differential Interference Contrast (DIC) Microscopy

    Design of a Transmitted Light Differential Interference Contrast Microscope

    Interpretation of a Transmitted Light Differential Interference Contrast Image

    Design of a Reflected Light Differential Interference Contrast Microscope

    Interpretation of a Reflected Light Differential Interference Contrast Image

    Chapter 10. Amplitude Modulation Contrast Microscopy

    Hoffman Modulation Contrast Microscopy

    Reflected Light Hoffman Modulation Contrast Microscopy

    The Single-Sideband Edge Enhancement Microscope

    Web Resources

    Chapter 11. Fluorescence Microscopy

    Discovery of Fluorescence

    Physics of Fluorescence

    Design of a Fluorescence Microscope

    Fluorescence Probes

    Pitfalls and Cures in Fluorescence Microscopy

    Web Resources

    Chapter 12. Various Types of Microscopes and Accessories

    Confocal Microscopes

    Total Internal Reflectance Fluorescence Microscopy

    Superresolution With Wide-Field Microscopes

    Fluorescence Resonance Energy Transfer

    Fluorescence Redistribution After Photobleaching

    Laser Microbeam Microscope

    Optical Tweezers

    Laser Capture Microdissection

    Laser Doppler Microscope

    Centrifuge Microscope

    X-Ray Microscope

    Infrared Microscope

    Nuclear Magnetic Resonance Imaging Microscope

    Stereo Microscopes

    Scanning Probe Microscopes

    Acoustic Microscope

    Horizontal and Traveling Microscopes

    Microscopes for Children

    Microscope Accessories

    Web Resources

    Chapter 13. Video and Digital Microscopy

    The Value of Video and Digital Microscopy

    Video and Digital Cameras: The Optical-to-Electrical Signal Converters

    Monitors: Conversion of an Electronic Signal into an Optical Signal

    Storage of Video and Digital Images

    Connecting a Video System

    Web Resources

    Chapter 14. Image Processing and Analysis

    Analog Image Processing

    Digital Image Processing

    Enhancement Functions of Digital Image Processors

    Analysis Functions of Digital Image Processors

    Toupview Digital Image Processing Software

    The Ethics of Digital Image Processing

    Web Resources

    Chapter 15. Laboratory Exercises

    Laboratory 1: The Nature of Light and Geometric Optics

    Laboratory 2: Physical Optics

    Laboratory 3: The Bright-Field Microscope and Image Formation

    Laboratory 4: Phase-Contrast Microscopy, Dark-Field Microscopy, Rheinberg Illumination, and Oblique Illumination

    Laboratory 5: Fluorescence Microscopy

    Laboratory 6: Polarized Light

    Laboratory 7: Polarizing Light Microscopy

    Laboratory 8: Interference Microscopy

    Laboratory 9: Differential Interference Contrast Microscopy and Hoffman Modulation Contrast Microscopy

    Laboratory 10: Video and Digital Microscopy and Analog and Digital Image Processing

    Web Resources


    Appendix I. Light Microscopy: A Retrospective

    Web Resource

    Appendix II. A Final Exam

    Appendix III. A Microscopist’s Model of the Photon


    Color Plates

Product details

  • No. of pages: 366
  • Language: English
  • Copyright: © Academic Press 2014
  • Published: December 16, 2013
  • Imprint: Academic Press
  • Hardcover ISBN: 9780124114845
  • eBook ISBN: 9780124115361

About the Author

Randy Wayne

Randy O. Wayne is a plant cell biologist at Cornell University notable for his work on plant development. In particular, along with his colleague Peter K. Hepler, Wayne established the powerful role of calcium in regulating plant growth; accordingly, their 1985 article, Calcium and plant development, was cited by at least 405 subsequent articles to earn the "Citation Classic" award from Current Contents magazine and has been cited by hundreds more since 1993. He is an authority on how plant cells sense gravity through pressure, on the water permeability of plant membranes, light microscopy, as well as the effects of calcium on plant development. He has published over 50 articles and is the author of another book, Light and Video Microscopy.

Affiliations and Expertise

Plant cell biologist at Cornell University

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