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

Contrast

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

References

Appendix I. Light Microscopy: A Retrospective

Web Resource

Appendix II. A Final Exam

Appendix III. A Microscopist’s Model of the Photon

Index

Color Plates