Page 12 - Fundamentals of Light Microscopy and Electronic Imaging

PREFACE

Throughout the writing of this book my goal has been how to teach the beginner how to
use microscopes. In thinking about a cover, my initial plan was to suggest a silhouette
of a microscope under the title “Practical Light Microscopy.” However, the needs of the
scientific community for a more comprehensive reference and the furious pace of elec-
tronic imaging technologies demanded something more. Practitioners of microscopy
have long required an instructional text to help align and use a microscope—one that
also reviews basic principles of the different optical modes and gives instructions on
how to match filters and fluorescent dyes, choose a camera, and acquire and print a
microscope image. Advances in science and technology have also profoundly changed
the face of light microscopy over the past ten years. Instead of microscope and film cam-
era, the light microscope is now commonly integrated with a CCD camera, computer,
software, and printer into electronic imaging systems. Therefore, to use a modern
research microscope, it is clear that research scientists need to know not only how to
align the microscope optics, but also how to acquire electronic images and perform
image processing. Thus, the focus of the book is on the integrated microscope system,
with foundations in optical theory but extensions into electronic imaging. Accordingly,
the cover shows the conjugate field and aperture planes of the light microscope under
the title “Fundamentals of Light Microscopy and Electronic Imaging.”
The book covers three areas: optical principles involved in diffraction and image
formation in the light microscope; the basic modes of light microscopy; and the compo-
nents of modern electronic imaging systems and the basic image-processing operations
that are required to prepare an image. Each chapter is introduced with theory regarding
the topic at hand, followed by descriptions of instrument alignment and image interpre-
tation. As a cell biologist and practitioner of microscopy rather than a physicist or devel-
oper of new microscope equipment and methods, the reader will notice that I have
focused on how to align and operate microscopes and cameras and have given somewhat
abbreviated treatment to the physical theory and principles involved. Nevertheless, the
theory is complete enough in its essentials that I hope even experienced microscopists
will benefit from many of the descriptions. With the beginner microscopist in mind,
each chapter includes practical demonstrations and exercises. The content, though not
difficult, is inherently intricate by nature, so the demonstrations are valuable aids in
absorbing essential optical principles. They also allow time to pause and reflect on the xi