Description

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.

Table of Contents

Series Page

Contributors

Preface

Chapter 1. Microscope Basics

Introduction

1.1 How Microscopes Work

1.2 Objective Basics

1.3 Mounting Video Cameras on the Microscope

Reference

Chapter 2. The Optics of Microscope Image Formation

Introduction

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

Conclusions

2.9 Appendix A

2.10 Appendix B

2.11 Appendix C

Acknowledgments

References

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

Acknowledgments

Chapter 4. Fundamentals of Fluorescence and Fluorescence Microscopy

Introduction

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

Acknowledgments

References

Chapter 5. Fluorescent Protein Applications in Microsc

Details

No. of pages:
672
Language:
English
Copyright:
© 2013
Published:
Imprint:
Academic Press
eBook ISBN:
9780124078925
Print ISBN:
9780124077614

About the editors

Greenfield Sluder

Affiliations and Expertise

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

David Wolf

Affiliations and Expertise

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