Digital Microscopy, Volume 114

4th Edition

Editors: Greenfield Sluder David Wolf
Hardcover ISBN: 9780124077614
eBook ISBN: 9780124078925
Imprint: Academic Press
Published Date: 18th August 2013
Page Count: 672
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Table of Contents

Series Page



Chapter 1. Microscope Basics


1.1 How Microscopes Work

1.2 Objective Basics

1.3 Mounting Video Cameras on the Microscope


Chapter 2. The Optics of Microscope Image Formation


2.1 Physical Optics: The Superposition of Waves

2.2 Huygens’ Principle

2.3 Young’s Experiment: Two-Slit Interference

2.4 Diffraction from a Single Slit

2.5 The Airy Disk and the Issue of Microscope Resolution

2.6 Fourier or Reciprocal Space: The Concept of Spatial Frequencies

2.7 Resolution of the Microscope

2.8 Resolution and Contrast


2.9 Appendix A

2.10 Appendix B

2.11 Appendix C



Chapter 3. Proper Alignment of the Microscope

3.1 Key Components and Image Locations of the Light Microscope

3.2 Working the Microscope: Basic Setup

3.3 Addendum


Chapter 4. Fundamentals of Fluorescence and Fluorescence Microscopy


4.1 Light Absorption and Beer’s Law

4.2 Atomic Fluorescence

4.3 Organic Molecular Fluorescence

4.4 Excited State Lifetime and Fluorescence Quantum Efficiency

4.5 Excited State Saturation

4.6 Nonradiative Decay Mechanisms

4.7 Fluorescence Resonance Energy

4.8 Fluorescence Depolarization

4.9 Measuring Fluorescence in the Steady State

4.10 Construction of a Monochromator

4.11 Construction of a Photomultiplier Tube

4.12 Measuring Fluorescence in the Time-Domain

4.13 Filters for the Selection of Wavelength

4.14 The Fluorescence Microscope

4.15 The Power of Fluorescence Microscopy



Chapter 5. Fluorescent Protein Applications in Microsc


The previous edition of this book marked the shift in technology from video to digital camera use with microscope use in biological science. This new edition presents some of the optical fundamentals needed to provide a quality image to the digital camera. Specifically, it covers the fundamental geometric optics of finite- and infinity-corrected microscopes, develops the concepts of physical optics and Abbe’s theory of image formation, presents the principles of Kohler illumination, and finally reviews the fundamentals of fluorescence and fluorescence microscopy. The second group of chapters deals with digital and video fundamentals: how digital and video cameras work, how to coordinate cameras with microscopes, how to deal with digital data, the fundamentals of image processing, and low light level cameras. The third group of chapters address some specialized areas of microscopy that allow sophisticated measurements of events in living cells that are below the optical limits of resolution.

Key Features

  • Expands coverage to include discussion of confocal microscopy not found in the previous edition
  • Includes "traps and pitfalls" as well as laboratory exercises to help illustrate methods


Cell and molecular biologists and researchers utilizing digitial microscopy techniques


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© Academic Press 2013
Academic Press
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About the Editors

Greenfield Sluder Editor

Affiliations and Expertise

University of Massachusetts Medical Center, Shrewsbury, U.S.A.

David Wolf Editor

Affiliations and Expertise

University of Massachusetts Medical Center, Shrewsbury, U.S.A.